American Journal of Molecular Biology, 2013, 3, 204-214 AJMB Published Online October 2013 (
The origin of biological information and programmed
protein synthesis
Dan Liu
John Curtin School of Medical Research, Australian National University, Acton, Australia
Received 22 July 2013; revised 15 August 2013; accepted 2 September 2013
Copyright © 2013 Dan Liu. This is an open access article distributed under the Creative Commons Attribution License, which per-
mits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Biological information is one of the most important
characteristics of life, and it enables life to evolve to
higher complexity and adapt to the environment by
mutation and natural selection. However, the origin
of this information recording and retrieval system
remains a mystery. To understand the origin of bio-
logical informa tion will lea d us to one step closer to un-
derstand the origin of life on earth. Biological infor-
mation is encoded in DNA and translated into protein
by the ribosome in all free living organisms. The in-
formation has to be translated into proteins to carry
out its biological functions, so the evolution of the
ribosome must be integrated with the development of
biological information. In this article, I propose that
the small ribosomal subunit evolved from a ribozyme
that acted as an RNA helicase in the ancient RNA
world, and the involvement of tRNAs and the large
ribosomal subunit evolved to enhance the helicase
activity and to overcome the higher energy require-
ment for high GC content RNA helices. This process
could have developed as a primitive recording mecha-
nism: since Watson-Crick base paring is a natural
property of RNA, each time the proto-small ribo-
somal subunit came to a particular GC-rich helix,
tRNA-like molecules and the proto-large ribosomal
subunit would have to be engaged to generate the he-
licase activity, and consequently the same polypep-
tide would be synthesized as a by-product. Simple re-
corded messages then evolved into useful biological
information through continuous mutation and natu-
ral selection. This hypothesis provides logical and in-
cremental steps for the development of programmed
protein synthesis. I also argue that the helicase activity
is preserved in the modern ribosome and that from
our knowledge of the ribosome, and we can deduce
the possible mechanisms of the helicase activity.
Keywords: Ribosome; tRNA; Translation; Translocation;
mRNA Helicase; Evolution; Origin of Biological
Biological information is contained in all the genes and
intergenic regions with signals for gene expression.
There are two major types of genes: one encodes infor-
mation that is expressed as functional RNA molecules
such as rRNA, tRNA, and snRNA, and another encodes
information for protein synthesis expressed as mRNA.
The functional RNA genes are thought to be remnants of
an ancient RNA world [1-5] that existed prior to DNA
and proteins, and their functions are involved in RNA
editing or protein synthesis. The origin of the protein
coding genes is unknown, and it is the major focus of
this communication. Protein is on e of the most important
basic building blocks for life, and proteins are synthe-
sized by the ribosome according to genetic information
encoded in the genes of all living organisms. Since a
gene can only carry out its biological function after it is
translated into protein by the ribosome, the understand-
ing of the evolution, structure and function of the ribo-
some should lead to an understanding of the origin of
genetic information.
The ribosome is a large ribonucleoprotein complex
and consists of small and large subunits (in bacteria 30S
and 50S subunits, respectively, Figure 1(a). Transfer
RNA (tRNA) is one of the key substrates in protein syn-
thesis, and has an L-shaped structure with an anticodon
at the end of one (anticodon) arm and a specific amino acid
is linked to the end of the other (acceptor) arm (Figure
1(b)). Within the small ribosomal subunit the anticodon
interacts with a codon on the message RNA (mRNA) by
Watson-Crick base pairing and this process is called
tRNA selection or decoding. When the cognate tRNA is
elected, it delivers a specific amino acid into the pepti- s
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214 205
Head forward
rotation Head backward
Head forward
rotation Head backward
Acceptor arm
Anticodon arm
Amino acid-ACC
Figure 1. The structure of E. coli 70S ribosome and the possible mRNA helicase f unction of the ribosome. (a) The structural
rearrangement of the 70S ribosome upon binding of mRNA and a P-site anticodon stem loop (ASL). Rearrangement of the 30S
head position in the apo-70S ribosome structure are indicated by different vectors between phosphorous atoms (light blue) and
atom (dark blue). In the ribosome complex, the 5’ to 3’ direction of mRNA is indicated, and letters A, P and E represent the
approximate positions of the tRNA binding sites at the subunit interface. Domains of the ribosome are labeled for the 30S head
(Head) and 50S central protuberance (CP), as are ribosomal protein in the small (S) and large (L) subunits (from Berk et al.
[37], Copyright (2006) National Academy of Sciences, U. S. A.) (b) The yeast tRNAPhe (PDB code 1EHZ) backbone structure
is shown with the last three nucleotides of the tRNA (C74, C75, and A76) labeled. The first half (5’) of the molecule is shown
in light blue and the second (3’) half in dark blue. (c) The two modes of helicase activity on the ribosome. Top panel (mode I)
the 30S subunit could move along mRNA with minimal structure in the 5’ to 3’ direction based on the head movement of the
30S subunit. Lower panel (mode II) when the small subunit engages with a high GC-content helix, it could stall, and tRNAs
could have a chance to bind to the mRNA and attract the large subunit. The movements of tRNAs in the ribosome could pro-
mote inter- and intra-subunit conformational changes, thereby enhancing helicase activity. The open arrows indicate the direc-
tion of the movement of the head domain and the large subunit and the acceptor part of tRNA are omitted in this illustration.
dyl transfer center (PTC) and a new peptidyl bond is
formed in the large subunit (Figure 2). In this way the
genetic information is converted into a specific amino
acid sequence in a protein. There are three tRNA binding
sites in both 30 S and 50 S ribosomal subunits (Figure 2):
the A site for the incoming aminoacyl-tRNA, the P site
for the peptidyl-tRNA, and the E site for the exiting dea-
cylated tRNA from the ribosome. After peptide bond
formation, the nascent peptide is transferred to the A site
bound tRNA, and the P site bound tRNA is deacylated
and spontaneously moves to the E site on the large sub-
unit, while the anticodon still binds on the P site on the
small subunit. This intermediate state with tRN As bound
to A/P and P/E positions is called the hybrid state (Fig-
ure 2) [6,7]. An elongation factor G (EF-G) catalyzes the
translocation of the mRNA/tRNA complex, and this
process moves both tRNAs from the A and P sites to the
P and E sites, respectively, and the mRNA advances by
one codon (see in reviews [8,9]). During protein synthe-
sis, the tRNA-mRNA complex is translocated through
the ribosome along a path of more then 100 Å, and the
translocation process involves a series of coordinated
conformational changes affecting both subunits.
Numerous attempts have been made to describe the
evolution of translation and the ribosome, such as Crick
[1], Woese [10], Noller [11], Poole [12], Fox and Naik
[13], and Wolf and Koonin [5] just to name a few. It has
been suggested that the two subunits evolved independ-
ently [11,14,15]. The evolution of the large subunit has
been subjected to intense studies and the suggestion has
been made that it possibly evolved earlier than the small
ubunit [13]. It was suggested that the large subunit s
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214
Hybrid state
Unlock ribosome
Hybrid state
Fully rotated
Rotate back
Relock ribosome
Amino aci
A, P, and E: the A, P,and E tRNA binding sites, respectivel
Helicase activity
and translocation
occur at this step
Figure 2. Elongation cycle of protein synthesis. The diagrams illustrate the inter-subunit rotation and the translocation
of tRNAs from the A and P sites to the P and E sites, respectively. After peptide-bond formation, the acceptor ends of
tRNAs spontaneously move from the A and P sites to the P and E sites, respectively, in the large ribosomal subunit
forming the hybrid state. EFG-GTP binds to the ribosome and hydrolyzes GTP to GDP, and stabilizes the rotation of
the small ribosomal subunit. At this step mRNA advances relative to the large subunit. When the small subunit rotates
back, the translocation of mRNA/tRNA complex relative to the small subunit occurs and the returning head of the
small subunit shears the mRNA helix at the entry of the downstream tunnel. This step is a novel suggestion in this
communication and is framed in a box. The open arrows indicate the direction of the movement of the 30S subunit and
its head domain.
could have evolved from a small RNA molecule of about
80 to110 nucleotides (nt) that could bind to tRNA-like
small RNA molecules on their 3’ CCA-amino acid ends.
When a duplication event occurred, the resulting mole-
cule could have formed two binding pockets side by side,
which eventually de veloped into the A and P sites. It has
been proposed that since two CCA-amino acid ends of
RNA molecules could bind simultaneously in a very
close proximity, it reduced the entropy and allowed pep-
tide bond formation and gave rise to the primitive pepti-
dyl transfer center (PTC) [13,16,17]. Recent studies [18,
19] seem to confirm this model. This reaction could have
led to the formatio n of oligopeptides with rando m amino
acid sequences. Random oligopeptides might offer
higher degrees of complexity for RNA structures and
capability for chemical catalysis [3,20] and could conse-
quently confer a huge advantage to the primitive “living”
system. Hence, production of random peptides was se-
lected. In order to improve the binding of the tRNA-like
small RNAs (or proto-tRNA, pt-tRNA) and improve the
efficiency of random oligopeptide synthesis driven by
the selection, the proto-ribosome could have expanded in
size and increased in complexity with evolutionary time.
However, the further evolution of the capacity to synthe-
size a specific protein was still not possible since at this
stage the peptide sequences were created by chance and
there was no recording mechanism for repeatable peptide
The origin of decoding must directly relate to the ori-
gin of biological information and programmed protein
synthesis. However, it is hard to imagine how the decod-
ing process or the coding system could have originated
independently, as the codes would not have any meaning
without a decoding system: a chicken-and-egg problem.
Since evolution has no foresight, the code assignment
and the decoding process had to evolve from other func-
tions within the RNA world. A recent study [21] has sug-
gested that the code assignment emerged before transla-
tion and could have been related to aminoacylating pri-
mordial tRNA by ribozymes mediated by the direct
stereochemical affinity between amino acids and anti-
codons (or codons) [22] possibly for genomic 3’ tag [14].
It has also been suggested that the small subunit possibly
evolved from RNA polymerase [12,23]. In this model,
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D. Liu / American Journal of Molecular Biology 3 (2013) 204-214 207
pt-tRNAs would interact with an mRNA-like template
(or proto-mRNA, pt-mRNA) in the proto-small subunit
(pt-small subunit), and in a fashion similar to that of the
codon and anticodon interaction. The “anticodon” was
then cleaved from the pt-tRNA molecule and lig ated with
the nascent RNA. However, there is no evidence to sug-
gest that the required nuclease or ligase activities ever
existed on the ribosome. Furthermore that hypothesis
does not provide a clear path that leads to programmed
protein synthesis, nor an explanation for the association
and coordinated actions of the small and large subunits.
Due to the redundancy of codon recognition, this type of
polymerization would have a mutation rate up to 30%
that is much greater than the minimum replication fidel-
ity (<1%) required to conserve genetic information [24,
25]. It was also suggested [12] that the pt-mRNA could
have served as an anchor for the pt-tRNAs to stabilize
them on the PTC, thereby incr easing the efficiency of the
reaction. The pt-small subunit would have evolved the
capacity to move the anchoring RNA in order to move
the pt-tRNA directionally out of the PTC after the reac-
tion. This hypothesis suggested that the anchoring RNA
evolved into information carrying molecule for protein
synthesis in the process [5,12,26]. However, this seems
unlikely since the available experimental data have
shown that the deacylated tRNA moves out of the PTC
before the translocation of the mRNA [7].
In this communication, I put forward an alternative
hypothesis that the small ribosomal subunit evolved from
an RNA helicase, and suggest that this function has been
preserved in the modern ribosome. In the RNA world, we
assume that the RNA had template-dependent replication,
but this would present a problem: how did the double-
stranded RNA separate? If the strands did not separate,
they could not form templates again or be folded into a
ribozyme and there would be no replication cycle. Possi-
bly at a very early stage, a geothermal pool could provide
the energy for strand separation and the molecules could
have circulated through the thermal gradient for their
replication cycles. However, when the pool cooled down
with geological changes over time, a helicase made from
RNA and for RNA would become critical and would be
as equally important as a polymerase under such condi-
tions. I propose that the pt-small subunit could have been
such a helicase. Furthermore pt-tRNA molecules could
have enhanced the helicase activity, especially for the
higher energy requirement of a GC-rich helix, by in-
creasing the grip of the pt-small subunit on the p t-mRNA
and by engaging the proto-ribosome (the large subunit).
These steps simultaneously produce a polypeptide as a
by-product. Since Watson-Crick base pairing is a natural
property of RNA, each time the pt-small subunit came to
the same GC-rich helix, the same polypeptide would
have been synthesized; in essence, the first oligonucleo-
tide reading mechanism linked to repeatable peptide
formation. At the very beginning, this recording system
would not have recorded any meaningful messages, but
through mutation and natural selection , messages associ-
ated with useful peptides were continuously enhanced
and conserved, giving rise to this type of biological in-
formation. This hypothesis is supported by the notion
that the tRNA aminoacylated with earliest amino acids,
such as glycine, alanine, valine and glutamic acid, which
could be synthesized abioticly under primitive earth con-
ditions [27], generally have the highest GC content in
their anticodon and have the most stable interaction with
the mRNA through Watson-Crick base pairing [28], and
this could be essential for the proposed helicase activity.
In fact, the coding regions in general have higher G + C
content compared to the non-coding regions even in
modern genes [29,30], which might be the traceable
mark of this process. A similar idea has been proposed by
Zenkin in 2012 [31], however, no possible mechanism of
the mRNA helicase activity related to the modern ribo-
some was given.
The mechanism proposed here can only work if the ri-
bosome has evolved from an mRNA helicase. Does any
trace of this activity remain on the modern ribosome?
The answer is affirmative. Noller and his colleagues have
shown that the ribosome itself is an mRNA helicase [32].
In a purified system that only contained E. coli ri-
bosomes, mRNA, tRNAs, and elongation factors EF-Tu
and EF-G, the ribosome was able to disrupt stably base-
paired RNA helixes, and the helicase active site was lo-
cated in the mRNA entry tunnel (downstream tunnel) at a
position +11 nt from the P site. In addition, helicase ac-
tivity has also been shown to occur in a system that had
only ribosomes, aa-tRNAs and the antibiotic sparsomy-
cin [32] that has previously been shown to promote
translocation without EF-G [6,33]. This indicates that the
helicase activity may be directly linked to translocation,
or that translocation is a p art of the helicase activity. The
coupling of helicase activity and translocation provides
an important clue to explain how the coding and decod-
ing system could have started, and shifts our fixed gaze
on protein synthesis as the sole function of the ribosome
to other possible functions.
A previous study using individual ribosomes and “op-
tical tweezer” techniques demonstrated that the ribosome
itself can unwind mRNA helices without an additional
mRNA helicase [34]. They suggested that translocation
and RNA unwinding are strictly coupled with ribosome
functions. A recent study indicated that the ribosome as
an mRNA helicase has two active mechanisms [35]. In
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214
the first mechanism, the mRNA helical junction was de-
stabilized at the entry site on the ribosome by the interac-
tions of the positively charged residues of the ribosomal
proteins (r-protein) S3 and S4 and the backbone of
mRNA at the helical junction. Mutation of these residues
reduced the helicase activity, but did not eliminate it [32].
In the second mechanism, the mRNA helix is mechani-
cally separated at the close junc tion during the inter- and
intra-subunit ribosomal conformational changes and this
was coupled with translocation. Qu and colleagues sug-
gested that the conformational changes “generate a force
that pulls on the tRNA: mRNA complex and promotes
unwinding at the mRNA entry site”. This mechanism
appears to be a unique property of the ribosomal mRNA
helicase activity [35]. They also found that the transla-
tion rate is greatly influenced by the G + C content of
folded structures in the mRNA at the ribosomal entry
These findings indicate that the modern ribosome is
still an mRNA helicase, and that the helicase activity is
directly linked to the inter- and intra-subunit conforma-
tional changes during an elongation cycle. Therefore it is
possible that this activity could have existed from the
beginning and could have been the primordial function
of the ancient ribosome before it acquired its function as
a programmed protein synthesis machine.
If the ribosome was and is an mRNA helicase, we should
be able to explain all of its functions in terms of helicase
activity. So, wh y is the decoding process (th e interaction
of tRNA and mRNA through Watson-Crick base paring)
important for helicase activity? The proper base pairing
of tRNA to mRNA increases the binding strength be-
tween the small subunit and the tRNA/mRNA complex
on the A site [36] and the P site [37]. This increases the
grip of the small subunit on mRNA. A mechanical study
[38] demonstrated that addition of an aa-tRNA analog
(N-acetylated Phe-tRNAphe) to an mRNA/ribosome com-
plex strengthened the mRNA-r ibosome bond.
In the current hypothesis, I su ggest that the additio n of
tRNA to the small subunit could attract the large subunit
and start the elongation cycle and the movements of
tRNAs in the ribosome promoting inter- and intra-sub-
unit conformational changes and therefore enhance the
helicase activity (Figure 1(c), lower panel). As previ-
ously shown [7], after peptidyl transfer, the acceptor end
of the deacylated tRNA travels spontaneously from the P
site to the E site in the large subunit and the tRNA binds
between the P/E positions (Figure 2). As a result of this
movement the L shaped tRNA acts like a spring with one
end bound to mRNA through the Watson-Crick base
pairing and the P site of the small subunit and the other
end interacts with the E site of the large subunits. The
distance between the P site and the E site in the large
subunit is about 50 Å [39,4 0], and the change in positio n
of tRNA from P/P to P/E creates a pulling force on the
small subunit, that strongly favors the rotated position of
the small subunit relative to the large subunit and the
rotation of the head of the small subunit. These actions
unlock the ribosome and are known to be important for
translocation [41], and are also associated with helicase
activity [35]. Improper base pairing of tRNA with mRNA
would weaken the mRNA/tRNA complex, affect trans-
location, and cause frameshift mutations [42,43]. If there
is no codon/anticodon interaction on the P site of the
small subunit, the tRNA could not properly engage with
the head of the small subunit and could not deliver the
pull from the E site of the large subunit to the mRNA and
the small subunit. Hence the interactions between the
mRNA and the small subunit could not be dislodged and
the translocation of mRNA would not occur. Conse-
quently, with codon/anticodon base pairing, tRNAs act
like a “handle” pulling mRNA forward, and this is why
decoding is important for helicase activity and how the
decoding mechanism could have been established before
the coding system even existed.
4.1. Helicase Mechanism Mode I
The ribosome has two possible modes of action as an
mRNA helicase. The first mode involves the action of the
small subunit without the involvement of tRNAs and th e
large subunit. It is well established that the head of the
small subunit is inherently dynamic in the absence of
binding of an anticodon stem loop (ASL) or a tRNA
(Figure 1(a), [37,44]). I propose that this characteristic
of the small subunit could mediate its helicase activity
and the small subunit could have been evolved from an
intrinsic RNA helicase ribozyme. It is likely that the
pt-mRNA bound to the P site of th e pt-small subunit in a
sequence-independent manner through the phosphori-
bose backbone. When the head rotated towards the E site,
the binding was no longer favorable in this position and
the pt-mRNA would have been released (Figure 1(c),
upper panel). When the head rotated back, a new interac-
tion could form again. The disruption of RNA base-
pairing possibly occurred during the head domain rota-
tion, or when the h ead rotates back away from th e E site.
Each cycle of translocation might have only disrupted
one base-pair (Figure 1(c), upper panel), since there
would be no reading frame.
This helicase activity is probably conserved in the
modern ribosome and may relate to the activity called the
ribosome scanning, when an mRNA is translo cating fro m
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214 209
5’ to 3’ without the engage ment of tRNAs. The scanning
activity of the ribosome can be observed at the initiation
of eukaryotic protein synthesis [45,46] or in mRNA with
translational bypass [47,48], and scanning is undertaken
by the small subunit alone [46] or the 70S ribosome [47].
A mechanical study of single ribosome/mRNA com-
plexes [38] showed that there were three distinct groups
of ribosome/mRNA complexes which were grouped ac-
cording to ruptur e force (needed to pull th e mRNA out of
ribosome) less than 6 pN, between 6 and 15 pN, and be-
tween 15 and 25 pN. It is possible to speculate that these
three groups represent the three different binding states
of mRNA on the ribosome: releasing, head rotating and
binding on the P site, respectively.
4.2. Helicase Mechanism Mode II
However, the proposed helicase activity of the (p t-)small
subunit alone could only work on pt-mRNAs that had
minimum secondary structure with a low G + C content,
as it would stall when it came to high G + C helices due
to their higher energy requirement. The prop osed second
mode of helicase activity evolved to bind the pt-aa-
tRNAs to the pt-mRNA/small subunit complex and pro-
mote its engagement with a proto-ribosome, and this
would have provided the extra energy for the helicase
activity to act on high G + C content mRNA helices as
discussed above (Figure 1(c), lower pa nel ). After peptid e
transf er, the hybrid sta te of tRNAs could pu ll the head of
the small subunit towards the E site. When the movement
is far enough and sustained long enough to release the
binding of the A and P sites of the small subunit to the
mRNA and the tRNAs, the spontaneous translocation
would occur. However, in an extant system this rarely
happens due to other forces acting on the mRNA/tRNAs
complex, such as the bending of h44, and the binding of
proteins S12 and S13 [37,49,50] that resist the rotation of
the head of the small subunit and pull the tRNAs in the
opposite direction back to the A/A and P/P positions. In
support of this, r ibosomes depleted of ribosomal p roteins
S12 and/or S13 increased spontaneous translocation [51],
indicating that the force pulling the head of the small
subunit towards the A site was reduced and thermody-
namically favored the rotated state and therefore increas-
ing spontaneous translocation. When ribosomes were
treated with the thiol-specific agent pCMB (p-chloro-
mercuribenzoate), spontaneous translocation occurred
and in some cases it could continue for greater then 40
elongation cycles [52]. The p-CMB agent targets the cys-
teine residues on the ribosomal proteins, indicating that
the r-proteins regulate inter- and intra-subunit move-
ments, and the regulatory action makes the movements
more precise and efficient and that is important for pro-
grammed protein synthesis. However, the initiation of
the movements and the movements themselves are most
likely induced by the interactions between tRNAs and
If translocation was an inherent process of a helicase,
in the very early stage in the development of the ribo-
some, it would not have had a strict reading frame. It is
likely that each step of translocation could disrupt mostly
3 base pairs of an RNA helix, but could be 2 or 4 base
pairs, depending on how far the small subunit’s head
could turn and how stable the RNA helices were; this
would not have impaired the function of a helicase. Only
when its by-products (proteins) were favorably selected
for their much higher catalytic ability and functional
versatility did a more measured translocation become
important in order to establish a reading frame to repro-
duce the same protein each time. This step was subse-
quently regulated by evolving proteins such as elonga-
tion factor G (EF-G) and other r-proteins. This process
would allow the development of programmed protein
synthesis by incremental steps.
It has been well established by the means of toeprinting
and the labeling of mRNA with pyrene [53-55] that when
the small subunit and its head are in the ro tated position,
the mRNA/tRNA complex is not yet translocated. This
indicates that the action of rotation do es not in itself pull
the mRNA in to the entry tunnel, an d means that the posi-
tion of mRNA relative to th e small subunit is unchanged.
However, relative to the large subunit, the mRNA/tRNA
complex is advanced by the rotation by at least 6 Å [56,
57]. A recent study showed conclusively that the mRNA
translocation occurred at the second step of inter-subunit
rotation [58]. The first step of translocation is the coun-
terclockwise rotation of the small subunit relative to the
large subunit that happens rapidly with or without EF-G
[59,60], after the P site tRNA moves to the hybrid state
conformation. The mechanism of this movement is not
yet known, but it was suggested that the distorted shape
of the P site tRNA might be the driving force for its
movement from the P/P to P/E position [50]. The anti-
codon arm of the P site tRNA is deformed by the oppos-
ing interaction with the head of the small subunit and
helix 69 of the large subunit; this results the opening of
the major groove at the 26:44 base pair of the anticodon
arm [50]. When the binding affinity between tRNA and
the P site of the large subunit is reduced after deacylation,
tRNA moves towards the E site to recover its more re-
laxed structure. Once the tRNAs move to the hybrid state,
it unlocks the ribosome and inter-subunit rotation can
take place. The second step of translocation is the clock-
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214
wise rotation that slowly restores the small subunit and
its head back to the non-rotated state, and this coincided
with the translocation of mRNA [58]. Furthermore, they
demonstrated that the second step does not require EF-G
release or GTP hydrolysis. It is possible that when the
contacts between the mRNA/tRNA complex and the
small subunit are finally broken, the torsion from the
twisted head of the small subunit and the inertia of the
reverse movement provide the disrupting force for the
helicase activity and the translocation. The mRNA entry
tunnel is very narrow at about 15 Å and only allows a
single strand of RNA to enter [61]. The returning head
shears the RNA helix on the entrance of the tunnel, and
pulls the mRNA into the tunnel, so the helicase activity
and the translocation happen all at once (Figure 2 boxed).
On the modern ribosome, the activities are enhanced by
S3 and S4 proteins [32]. While the head of the small
subunit is turning back, the mRNA is stab ilized solely by
the codon-antico don interaction with tRNAs on the P site
and E site. The tRNAs are in turn stabilized by the bind-
ing to the P site and the E site, and the L1 stalk on the
large subunit, and EF-G + GDP which leans against the P
site peptidyl-tRNA on the small subunit (Figure 2) [62,
63]. There will be 4 to 6 bases of codon-an ticodon inter-
action due to the redundancy and only 0 to 3 base-pairing
will need to be disrupted depending on the structure of
mRNA for each step of translocation, indicating that this
process is thermodynamically feasible. The current
model implies that there is an energy barrier (disrupting
an mRNA helix ) to overcome when the head of the small
subunit rotates back, and this process would put a strain
on mRNA/tRNA complex. Consideration of this model
provides a molecular explanation and demonstrates why
the codon/anticodon interactions in the P and/or E sites
are crucial for maintaining the reading frame. When
there are defects in codon-anticodon interaction on the P
site and/or the E site, it weakens the mRNA/tRNA com-
plex, and when the head of the small subunit is turning
towards the A site (the second step of translocation) and
disrupting the mRNA helix, the pull would dislodge the
mRNA from the tRNAs and cause frame shifts. This was
observed by Zaher and Green [43] in a recent study. In
some cases of programmed frameshifting, the pseudok-
not on the mRNA, which interacts with the ribosome at
the entrance of the downstream tunnel, presented a
higher energy barrier for the helicase activity and trans-
location. The returning head of the small subunit pushed
on the pseudoknot, and this dislodges mRNA from the
tRNAs and causes frame shifting [64]. Cryo-EM studies
on ribosomal intermediates stalled by a pseudoknot re-
vealed the strain on the mRNA, the bending of the tip of
h44 towards the A site, and distor tion of the P site tRNA
[64]. This observation suppo rts the current model. At the
other extreme, when the mRNA is devoid of secondary
structure, such as poly-U, the codon-anticodon interac-
tion was enough to stab ilize mRNA without EF-G during
translocation [65-67]. On the other hand, translocation
was severely inhibited even in the presence of EF-G +
GTP when the binding of tRNA in the P/E position was
affected by the introduction of mutations in the E site of
the large subunit [68,69], or on the CCA end of the P/E
bound tRNA [70-72], or in the absence of L1 protein [73].
This evidence indicates that tRNA is not only an adaptor
for protein synthesis, but also plays an essential role in
translocation by promoting ribosomal conformational
changes and this is in the heart of h e licase activity.
To summarize the possible of mechanism of helicase
activity and translocation: the hybrid state of tRNAs
strongly favors the rotation of the small subunit relative
to the large subunit and the rotation of its head towards
the E site. This movement partially advances the mRNA
relative to the large subunit. The rotation weakens the
binding between the small subunit and the mRNA/tRNA
complex [74], and eventually when the complex is re-
leased the tRNAs adopt the P and E site positions be-
cause of the binding of the large subunits to the tRNAs
on the P and E sites, respectively. At this stage, the
mRNA is completely translocated relative to the large
subunit, but the small subunit and its head are still rotat-
ing back. When the rotations are completed, the translo-
cation relative to the small subunit and helicase activity
are accomplished (Figure 2). In this process, most of the
contacts and movements are between rRNAs and tRNAs,
so one could envisage that the similar action could hap-
pen in an all RNA ribosome; albeit it might not be very
The origins of genetic information and programmed pro-
tein synthesis have been an enduring enigma for biolo-
gists since the discovery of DNA and the ribosome in the
1950s. The debate between “genetic heredity-first” and
“metabolism-first” still continues. The recent “protein
centric” views of evolution of life [75-77] suggested that
proteins evolved first before the ribosome, and that the
peptides or proteins could be synthesized by other pep-
tides or proteins in the system and this process was
“autocatalytic”. The hypothesis was based on phyloge-
netic studies of protein structures and RNA, and one of
main arguments was that the proteins with the oldest
folds were metabolic enzymes, instead of proteins related
to translation or RNA binding. The major difficulties of
this hypothesis are not only the lack of evidence for the
“autocatalytic” process, but also it is without a viable
mechanism to transfer the information of these proteins
to RNA or DNA. All the information of these ancient
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214 211
proteins would be lost in the transition from the noncod-
ing to the coding system and it is unlikely that the pro-
teins reinvented by coding synthesis would be identical
to their ancient counterparts, indicating that the methods
used in their study cannot reach the point beyond transla-
The present hypothesis describes how genetic infor-
mation could have been generated from the ancient RNA
world through a recording system and natural selection.
At the very early stage, the pt-small and pt-large subunits
as ribozymes could have evolved separately with differ-
ent functions, such as helicase and the production of
random peptides, respectively. Template-dependent rep-
lication and folded RNA structures form double helices
as a natural property of RNA, however these structures
could prevent further replication. To facilitate the essen-
tial helicase activity, the two subunits combined with
aminoacylated primordial tRNA could have cooperated
and produced random peptides as by-product. Some of
these peptides could have improved the fitness of the
primitive “living” system, including the ones that could
have basic metabolic fu nction or ability to stab ilize RNA
molecules. The RNA molecules encoding these peptides
would be selected and could form the basis for repeatable
peptide production. Mutations on these RNA molecules
could provide variants for natural selection and could
lead to a better adaptation to the environment for early
life. In this way, the genetic information could have
formed and continued to improve and diversify, and pave
the way for the transition from the RNA world to the
RNA/protein world. Th e ribosome could also mature and
become more efficient and have higher fidelity with the
involvement of proteins in the same process. Unlike the
previous models for the origin of the small ribosomal
subunit mentioned in introduction, which are sp eculative
and without experimental support, the present hypothesis
is based on the natural property of RNA molecules, the
principle of Darwinian evolution, and our current knowl-
edge of the ribosome and provides a sensible and logical
solution for the enduring enigma.
So far, I described the ribosome as an mRNA helicase
with two modes of action and reviewed the eviden ce that
the modern ribosome has the mRNA helicase function. I
believe that the helicase function of the ribosome existed
prior to and led to the emergence of programmed protein
synthesis. tRNA plays a central role in function in heli-
case activity. In 1970, Woese proposed that mRNA was
pulled through the ribosome by a tRNA ratchet [10]. Al-
though the detail of the proposed mechanism might not
be in agreement with our current knowledge of the ribo-
some, the basic idea was correct. Woese stated that “it
seems impossible to avoid invoking tRNA-like entities
(that is an ‘adaptor ’ system) as an integral part of transla-
tion from its very inception—which is sufficient reason
for suspecting the basic molecular mechanisms of the
process to lie in the properties of this molecular species”.
The present model describes the early inception of the
translation system and the role of tRNA based on the
properties of tRNA and our current knowledge of the
ribosome. This model also explains the necessity of the
coordinated movements of the small and large subunits,
and the possibility of the establishment of a recording
mechanism. The biological information was developed
from this recoding mechanism through continuous muta-
tion and natural selection. The interaction of the small
and large subunits could also improve the efficiency of
peptide bond formation, since the substrates (tRNAs) are
securely enveloped in the interface between the two
subunits, so the present proposal does not contradict
Fox’s large subunit-first model. It seems that the ribo-
some is the only example of this type of RNA helicase
existing in the modern world, and it is possible this heli-
case only evolved once in the history of evolution or
other similar RNA helicases were replaced by the most
successful pt-ribosome. It is still not clear which part of
the small subunit is original. It has been suggested that
the 3’ domain of the 16S rRNA including h44 and the P
site are the oldest parts of the small subunit [26,77-79],
and this is consistent with the present proposal. The dy-
namic movement of the head of the small subunit is at
the center of its helicase activity. Understanding the mo-
lecular basis that sustains this movement would lead to
the understanding of evolution of the small subunit.
It is widely argued that the RNA world preceded the
DNA and protein world. Today, although almost all the
functions that were carried out by RNAs in the RNA
world have been replaced by proteins, the function of the
ribosome, which is essentially RNA machinery, has not.
Proteins have not yet evolved the ability to synthesize
themselves according to genetic information after more
than three billion years of evolution. This gives us a
unique opportunity to glance back to the lost RNA world
through the structure and functions of the ribosome.
I thank Dr. David A. Bastin and Prof. Philip Board for their careful
reading of the manuscript and very helpful discussions, and Dr. Huina
Zhou for creating figures. I would like to thank Prof. Harry Noller for
his work on the ribosome and translocation that makes this work possi-
[1] Crick, F.R. (1968) The origin of genetic code. Journal of
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214
Molec ul ar Biology, 38, 367-379.
[2] Gilbert, W. (1986) The RNA world. Nature, 319, 618.
[3] Noller, H. (2004) The driving force for molecular evolu-
tion of transloation. RNA, 10, 1833-1837.
[4] Orgel, L.E. (1968) Evolution of the genetic apparatus.
Journal of Molecular Biology, 38, 381-393.
[5] Wolf, Y.I. and Koonin, E.V. (2007) On the origin of the
translation system and genetic code in the RNA world by
means of natural selection, exaptation, and subfunction-
alisztion. Biology Direct, 2, 14-39.
[6] Dorner, S., Brunelle, J.L., Sharma, D. and Green, R.
(2006) The hybrid state of tRNA binding is an authentic
translation elongation intermediate. Nature Structural &
Molecular Biology, 13, 234-241.
[7] Moazed, D. and Noller, H.F. (1989) Intermediate states in
the movement of transfer RNA in the ribosome. Nature,
342, 142-148.
[8] Joseph, S. (2003) After the ribosome structure: How does
translocation work? RNA, 9, 160-164.
[9] Korostelev, A., Ermolenko, D.N. and Noller, H.F. (2008)
Structural dynamics of the ribosome. Current Opinion in
Chemical Biology, 12, 674-683.
[10] Woese, C. (1970) Molecular mechanics of translation: A
reciprocating ratchet mechanism. Nature, 226, 817-820.
[11] Noller, H.F. (1993) On the origin of the ribosome: Co-
evolution of subdomains of tRNA and rRNA. In: Geste-
land, R.F. and Atkins, J.F. Eds., The RNA world, Cold
Spring Harbor Laboratory Press, Cold Spring Harbor,
New York, 137-156.
[12] Poole, A.M., Jeffares, D.C. and Penny, D. (1998) The
path from the RNA world. Journal of Molecular Evolu-
tion, 46, 1-17.
[13] Fox, G.E. and Naik, A.K. (2004) The evolutionary his-
tory of the translation machinery. In: De Pouplana, R.L.
Ed., The Genetic Code and The Origin of Life, Landers
BioScience, Georgetown, 92-105.
[14] Maizels, N. and Weiner, A.M. (1993) The genomic tag
hypothesis: Modern viruses as molecular fossils of an-
cient strategies for genomic replication. In: Gesteland,
R.F. and Atkins, J.F., Eds., The RNA world, Cold Spring
Harbor Laboratory Press, Cold Spring Harbor, New York,
[15] Schimmel, P., Giege, R., Moras, D. and Yokoyama, S.
(1993) An operational RNA code for amino acids and
possible relationship to genetic code. Proceedings of the
National Academy of Sciences of the United States of
America, 90, 8763-8768.
[16] Agmon, I., Bashan, A. and Yonath, A. (2006) On ribo-
some conservation and evolution. Israel Journal of Ecol-
ogy and Evolution, 52, 359-379.
[17] Sievers, A., Beringer, M., Rodnina, M.V. and Wolfenden,
R. (2004) The ribosome as an entropy trap. Proceedings
of the National Academy of Sciences of the United States
of America, 101, 7897-7901.
[18] Bokov, K. and Steinberg, S.V. (2009) A hierarchical mo-
del for evolution of 23S ribosomal RNA. Nature, 457,
[19] Hsiao, C., Mohan, S., Kalahar, B.K. and Williams, L.D.
(2009) Peeling the onion: ribosomes are ancient molecu-
lar fossils. Molecular Biology and Evolution, 26, 2415-
[20] Mzathmary, E. and Smith, M. (1997) From replicators to
re-producers: The first major transition leading to life.
Journal of Theoretical Biology, 187, 555-571.
[21] Rodin, A.S., Szathmary, E. and Rodin, S.N. (2011) On
origin of genetic code and tRNA before translation. Bi-
ology Direct, 6, 14.
[22] Rodin, A.S., Szathmary, E. and Rodin, S.N. (2009) On
ancestor for codes viewed from the perspective of two
complementary modes of tRNA aminoacylation. Biology
Direct, 4, 4.
[23] Gordon, K.H.J. (1995) Were RNA replication and trans-
lation directly coupled in the RNA (+protein?) world?
Journal of Theoretical Biology, 173, 179-193.
[24] Eigen, M. (1971) Selforganization of matter and the evo-
lution of biological macromolecules. Die Naturwissen-
schaften, 58, 465-523.
[25] Kun, A., Santos, M. and Szathmary, E. (2005) Real ri-
bozymes suggest a relaxed error threshold. Nature Ge-
netics, 37, 1008-1011.
[26] Fox, G.E. (2010) Origin and evolution of the ribosome.
Cold Spring Harbor Perspectives in Biology, 9, a003483.
[27] Miller, S.L. (1953) A production of amino acids under
possible primitive earth conditions. Science, 117, 528-
[28] Travers, A. (2006) The evolution of the genetic code revi-
sited. Origins of Life and Evolution of Biospheres, 36,
[29] Bultrini, E. and Pizzi, E. (2006) A new parameter to
study compositional properties of non-coding regions in
eukaryotic genomes. Gene, 385, 75-82.
[30] Grantham, R., Gautier, C., Gouy, M., Mercier, R. and
Pave, A. (1980) Codon catalog usage and the genome
hypothesis. Nucleic Acids Research, 8, r49-r62.
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214 213
[31] Zenkin, N. (2012) Hypothesis: Emergence of translation
as a result of RNA helicase evolution. Journal of Mo-
lecular Evolution, 74, 249-256.
[32] Takyar, S., Hickerson, R.P. and Noller, H.F. (2005) mRNA
helicase activity of the ribosome. Cell, 120, 49-58.
[33] Fredrick, K. and Noller, H.F. (2003) Catalysis of ribo-
somal translocation by sparsomycin. Science, 300, 1159-
[34] Wen, J.D., Lancaster, L., Hodges, C., Zeri, A.C., Yoshi-
mura, S.H. and Noller, H.F., et al. (2008) Following
translation by single ribosomes one codon at a time. Na-
ture, 452, 598-603.
[35] Qu, X., Wen, J.D., Lancaster, L., Noller, H.F., Busta-
mante, C. and Tinoco, I. Jr. (2011) The ribosome uses
two active mechanisms to unwind messenger RNA dur-
ing translation. Nature, 475, 118-121.
[36] Ogle, J.M., Brodersen, D.E., Clemons Jr., W.M., Tarry,
M.J., Carter, A.P. and Ramakrishnan, V. (2001) Recogni-
tion of cognate transfer RNA by the 30S ribosomal sub-
unit. Science, 292, 897-902.
[37] Berk, V., Zhang, W., Pai, R.D. and Cate, J.H. (2006)
Structural basis for mRNA and tRNA positioning on the
ribosome. Proceedings of the National Academy of Sci-
ences of the United States of America, 103, 15830-15834.
[38] Vanzi, F., Takagi, Y., Shuman, H., Cooperman, B.S. and
Goldman, Y.E. (2005) Mechanical studies of single ribo-
some/mRNA complexes. Biophysical Journal, 89, 1909-
[39] Korostelev, A., Trakhanov, S., Laurberg, M. and Noller,
H.F. (2006) Crystal structure of a 70S ribosome-tRNA
complex reveals functional interactions and rearrange-
ments. Cell, 126, 1065-1077.
[40] Noller, H.F. and Baucom, A. (2002) Structure of the 70 S
ribosome: Implications for movement. Biochemical Soci-
ety Transactions, 30, 1159-1161.
[41] Horan, L.H. and Noller, H.F. (2007) Intersubunit move-
ment is required for ribosomal translocation. Proceedings
of the National Academy of Sciences of the United States
of America, 104, 4881-4885.
[42] Muldoon-Jacobs, K.L. and Dinman, J.D. (2006) Specific
effects of ribosome-tethered molecular chaperones on
programmed -1 ribosomal frameshifting. Eukaryotic Cell,
5, 762-770.
[43] Zaher, H.S. and Green, R. (2009) Quality control by the
ribosome following peptide bond formation. Nature, 457,
[44] Schuwirth, B.S., Borovinskaya, M.A., Hau, C.W., Zhang,
W., Vila-Sanjurjo, A., Holton, J.M., et al. (2005) Struc-
tures of the bacterial ribosome at 3.5 A resolution. Sci-
ence, 310, 827-834.
[45] Kozak, M. (1978) How do eucaryotic ribosomes select
initiation regions in messenger RNA? Cell, 15, 1109-
[46] Kozak, M. (1989) The scanning model for translation: An
update. The Journal of Cell Biology, 108, 229-241.
[47] Herr, A.J., Atkins, J.F. and Gesteland, R.F. (1999) Muta-
tions which alter the elbow region of tRNA2Gly reduce
T4 gene 60 t ran sl at i ona l by pa ssi ng e f fic ien cy . The EMBO
Journal, 18, 2886-2896.
[48] Weiss, R.B., Huang, W.M. and Dunn, D.M. (1990) A
nascent peptide is required for ribosomal bypass of the
coding gap in bacteriophage T4 gene 60. Cell, 62, 117-
[49] Ogle, J.M., Murphy, F.V., Tarry, M.J. and Ramakrishnan,
V. (2002) Selection of tRNA by the ribosome requires a
transition from an open to a closed form. Cell, 11 1, 721-
[50] Selmer, M., Dunham, C.M., Murphy, F.V., Weixlbaumer,
A., Petry, S., Kelley, A.C., et al. (2006) Structure of the
70S ribosome complexed with mRNA and tRNA. Sci-
ence, 313, 1935-1942.
[51] Cukras, A.R., Southworth, D.R., Brunelle, J.L., Culver,
G.M. and Green, R. (2003) Ribosomal proteins S12 and
S13 function as control elements for translocation of the
mRNA: tRNA complex. Molecular Cell, 12, 321-328.
[52] Southworth, D.R., Brunelle, J.L. and Green, R. (2002)
EFG-independent translocation of the mRNA: tRNA
complex is promoted by modification of the ribosome
with thiol-specific reagents. Journal of Molecular Biol-
ogy, 324, 611-623.
[53] Pan, D., Kirillov, S.V. and Cooperman, B.S. (2007) Ki-
netically competent intermediates in the translocation
step of protein synthesis. Molecular Cell, 25, 519-529.
[54] Spiegel, P.C., Ermolenko, D.N. and Noller, H.F. (2007)
Elongation factor G stabilizes the hybrid-state conforma-
tion of the 70S ribosome. RNA, 13, 1473-1482.
[55] Zavialov, A.V., Hauryliuk, V.V. and Ehrenberg, M.
(2005) Guanine-nucleotide exchange on ribosome-bound
elongation factor G initiates the translocation of tRNAs.
Journal of Biology, 4, 9.
[56] Agirrezabala, X., Lei, J., Brunelle, J.L., Ortiz-Meoz, R.F.,
Green, R. and Frank, J. (2008) Visualization of the hybrid
state of tRNA binding promoted by spontaneous ratchet-
ing of the ribosome. Molecular Cell, 32, 190-197.
[57] Dunkle, J.A., Wang, L., Feldman, M.B., Pulk, A., Chen,
V.B., Kapral, G.J., et al. (2011) Structures of the bacterial
Copyright © 2013 SciRes. OPEN ACCESS
D. Liu / American Journal of Molecular Biology 3 (2013) 204-214
Copyright © 2013 SciRes.
ribosome in classical and hybrid states of tRNA binding.
Science, 332, 981-984.
[58] Ermolenko, D.N. and Noller, H.F. (2011) mRNA trans-
location occurs during the second step of ribosomal in-
tersubunit rotation. Nature Structural & Molecular Biol-
ogy, 18, 457-462.
[59] Cornish, P.V., Ermolenko, D.N., Noller, H.F. and Ha, T.
(2008) Spontaneous intersubunit rotation in single ri-
bosomes. Molecular Cell, 30, 578-588.
[60] Fischer, N., Konevega, A.L., Wintermeyer, W., Rodnina,
M.V. and Stark, H. (2010) Ribosome dynamics and
tRNA movement by time-resolved electron cryomicro-
scopy. Nature, 466 , 329-333.
[61] Borovinskaya, M.A., Shoji, S., Holton, J.M., Fred rick, K.
and Cate, J.H.D. (2007) A steric block in translation
caused by the antibiotic spectinomycin. ACS Chemical
Biology, 2, 545-552.
[62] Frank, J. and Agrawal, R.K. (2001) Ratchet-like move-
ments between the two ribosomal subunits: Their impli-
cations in elongation factor recognition and tRNA trans-
location. Cold Spring Harbor Symposia on Quantitative
Biology, 66, 67-75.
[63] Julian, P., Konevega, A.L., Scheres, S.H., Lazaro, M., Gil,
D., Wintermeyer, W., et al. (2008) Structure of ratcheted
ribosomes with tRNAs in hybrid states. Proceedings of
the National Academy of Sciences of the United States of
America, 105, 16924-16927.
[64] Namy, O., Moran, S.J., Stuart, D.I., Gilbert, R.J. and
Brierley, I. (2006) A mechanical explanation of RNA
pseudoknot function in programmed ribosomal frame-
shifting. Nature, 441, 244-247.
[65] Gavrilova, L.P., Kostiashkina, O.E., Koteliansky, V.E.,
Rutkevitch, N.M. and Spirin, A.S. (1976) Factor-free
(“non-enzymic”) and factor-dependent systems of trans-
lation of polyuridylic acid by Escherichia coli ribosomes.
Journal of Molecular Biology, 101, 537-552.
[66] Gavrilova, L.P. and Spirin, A.S. (1971) Stimulation of
“non-enzymic” translocation in ribosomes by p-chloro-
mercuribenzoate. FEBS Letters, 17, 324-326.
[67] Pestka, S. (1974) Assay for nonenzymatic and enzymatic
translocation with Escherichia coli ribosomes. Methods
in Enzymology, 30, 462-470.
[68] Sergiev, P.V., Lesnyak, D.V., Kiparisov, S.V., Bu-
rakovsky, D.E., Leonov, A.A., Bogdanov, A.A., et al.
(2005) Function of the ribosomal E-site: A mutagenesis
study. Nucleic Acids Research, 33, 6048-6056.
[69] Walker, S.E., Shoji, S., Pan, D., Cooperman, B.S. and
Fredrick, K. (2008) Role of hybrid tRNA-binding states
in ribosomal translocation. Proceedings of the National
Academy of Sciences of the United States of America, 105,
[70] Feinberg, J.S. and Joseph, S. (2001) Identification of
molecular interactions between P-site tRNA and the ri-
bosome essential for translocation. Proceedings of the
National Academy of Sciences of the United States of
America, 98, 11120-11125.
[71] Lill, R., Robertson, J.M. and Wintermeyer, W. (1989)
Binding of the 3’ terminus of tRNA to 23S rRNA in the
ribosomal exit site actively promotes translocation. The
EMBO Journal, 8, 3933-3938.
[72] Virumae, K., Saarma, U., Horowitz, J. and Remme, J.
(2002) Functional importance of the 3’-terminal adeno-
sine of tRNA in ribosomal translation. The Journal of
Biological Chemistry, 277, 24128-24134.
[73] Subramanian, A.R. and Dabbs, E.R. (1980) Functional
studies on ribosomes lacking protein L1 from mutant Es-
cherichia coli. European Journal of Biochemistry, 112,
[74] Uemura, S., Dorywalska, M., Lee, T.H., Kim, H.D., Pug-
lisi, J.D. and Chu, S. (2007) Peptide bond formation de-
stabilizes shine-dalgarno interaction on the ribosome.
Nature, 446, 454-457.
[75] Caetano-Anolles, D., Kim, K.M., Mittenthal, J.E. and
Caetano-Anolles, G. (2011) Proteome evolution and the
metabolic origins of translation and cellular life. Journal
of Molecular Evolution, 72, 14-33.
[76] Caetano-Anolles, G., Kim, K.M. and Caetano-Anolles, D.
(2012) The phylogenomic roots of modern biochemistry:
Origins of proteins, cofactors and protein biosynthesis.
Journal of Molecular Evolution, 74, 1-34.
[77] Harish, A. and Caetano-Anolles, G. (2012) Ribosomal
history reveals origins of modern protein synthesis. PloS
One, 7, e32776.
[78] Bernhardt, H.S. and Tate, W.P. (2010) The transition
from noncoded to coded protein synthesis: Did coding
mRNAs arise from stability-enhancing binding partners
to tRNA? Biology Direct, 5, 16-29.
[79] Schafer, M.A., Tastan, A.O., Patzke, S., Blaha, G., Spahn,
C.M., Wilson, D.N., et al. (2002) Codon-anticodon inter-
action at the P site is a prerequisite for tRNA interaction
with the small ribosomal subunit. The Journal of Bio-
logical Chemistry, 277, 19095-19105.