REVIEW

Re-creating the RNA world

Ichiro Hirao and Andrew D. Ellington
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.

Results from in vitro selection experiments can be used to construct and

test models for the evolution of the RNA world. Surprisingly, the success of
selected RNAs at binding ligands and catalyzing reactions may make it
difficult to determine precisely the lineage of molecular fossils, molecules
that are believed to have survived from the RNA world to the present.

Introduction allowed models of molecular evolution to be recapitu-

The discovery of catalytic RNA has dramatically shaped
speculations about the origin of life and the evolution of
metabolism. The primordial 'chicken and egg' conun-
drum - which came first, informational polynucleotides
or functional polypeptides? - was obviated by the
simple but elegant compaction of both genetic informa-
tion and catalytic function into the same molecule.
Moreover, the chemical complexity of catalytic RNAs
(ribozymes) led to speculations that the biochemistry of
the earliest organisms may have been a consequence of
the shapes and functions that nucleic acids could assume.
In the most intricate scenarios, the earliest RNA replica-
tors may have directly emerged from the 'prebiotic soup'
and subsequently evolved to become an 'RNA world'
[1], in which primordial biochemistry and metabolism
mirrored that found in modern life [2].
The conceptual revolution in our understanding of
nucleic acid evolution has been paralleled by an equally
profound technical revolution. Techniques such as DNA
sequencing, in vitro transcription and the polymerase
chain reaction have made it possible to manipulate and
characterize nucleic acid sequences almost completely ex
vivo. In turn, the principles that govern the natural selec-
tion of organisms can now be applied to molecules in a
test tube: a pool of heritably variable nucleic acid mol-
ecules can be prepared and sieved for sequences that
confer an improved phenotype, these phenotypes can be
allowed to compete among themselves, and the survivors
can be preferentially amplified. Although the notion of
in vitro selection is sometimes claimed to have itself arisen
de novo [3], it is a logical and obvious consequence of a
body of research into molecular evolution [4,5]: it was
the available techniques that advanced to the point where
old ideas became more experimentally accessible.
Models for molecular evolution
The intersection of the discovery of ribozymes with the
development of techniques for nucleic acid amplification
lated in a test tube. As extant catalytic RNAs are either
analogous to, or descended from, early ribozymes, many
of the ideas advanced under the general aegis of the
'RNA world' hypothesis can be subjected to experimen-
tal tests. Both the data gathered and their interpretation
can be scrutinized to determine whether a particular
aspect of a particular model for the RNA world might
have occurred, or whether an alternative scenario was
more likely. But although the data garnered from in vitro
selection experiments can be used to guide model-build-
ing, care must still be taken to ensure that the models
themselves are compatible with other facts and logically
self-consistent.
As a guide to how primordial molecular biology can
potentially be modelled by experiment and inference, we
will examine three claims that have been advanced
regarding different aspects of the 'RNA world' hypo-
thesis: that RNA catalysts can be functionally diverse,
that the origin of the genetic code may lie in amino
acid-RNA interactions, and that the evolution of the
translation apparatus and other functional RNAs could
have been guided by interactions with low molecular
weight effectors. By comparing and contrasting how evi-
dence is used to support or contradict these claims, it
should be possible to gain an understanding of the value
and limitations of using artificial evolutionary experi-
ments today to describe natural evolutionary processes
that may have occurred in the past.
The RNA world and the diversity of RNA
catalysts
In order to establish a common ground for discussion, a
generalized view of the RNA world is shown in Figure 1.
In this model, the earliest self-replicating nucleic acids -
whether RNA, DNA or some analogous polymer -
were elaborated into all-RNA genomes and catalysts. The
survival of virtually any self-replicating system is depen-
dent on avoiding dissipation by diffusion, and it is
expected that cellularization was one of the earliest

© Current Biology 1995, Vol 5 No 9 1017

1018 Current Biology 1995, Vol 5 No 9
phenotypic properties of a ribo-organism. DNA was
found to be a superior macromolecule for storing genetic
information, and proteins were a fortuitous side-product
of a complex ribozyme that could catalyze template-
directed peptide-bond formation. The invention of the
translation apparatus was likely to have been a cataclysmic
event that allowed superior protein catalysts to displace
biochemically complex, but less efficient, ribozymes.
Most of these evolutionary events occurred long before
the last common ancestor of modern life, the progenote,
gave rise to the three modern domains (eubacteria,
archaebacteria, and eukaryotes). Some vestiges of RNA's
former greatness can, however, still be found in (catalyti-
cally competent?) ribosomal RNA and its attendant trans-
fer RNAs, the nucleotide 'signatures' of ubiquitous cofac-
tors such as NAD and ATP, and perhaps in idiosyncratic
molecular fossils such as the group I self-splicing intron.
This scenario makes a strong prediction regarding the cat-
alytic functions that can be assumed by ribozymes. For
example, if NAD is a 'ribo-cofactor' descended from the
RNA world, then it should be possible for at least some
RNA sequences to act as dehydrogenases [6]. Similarly,
the fact that the cellular currency for energy, ATP, is a
nucleotide suggests that there may have been enzymes
such as ribo-kinases. In fact, the RNA world hypothesis
necessarily assumes the existence of a wide variety of ribo-
zymes, with functions that would have spanned the bio-
chemical reactions of intermediary metabolism. If these
Fig. 1. A generalized view of the RNA
world. Events hypothesized to have
occurred between the origin of life and
the progenote (the last common ances-
tor of modern life) are roughly ordered.
The chronology flows from 'early' at the
top of the page (- 3.5 billion years ago)
to 'late' at the bottom. Cellularization
represents the immobilization of self-
replicators, and could as easily have
occurred on a two-dimensional surface
as within a lipid vesicle. As diffusion
would have severely reduced the fitness
of replicators, cellularization may have
been concomitant with the origin of life.
Similarly, as the resupply of metabolites
from the soup would probably have
superseded the need for an error-resis-
tant genome, metabolism is shown to
occur before the invention of DNA.
'Molecular fossils' that may have per-
sisted from earlier versions of life are
represented on the right.
catalytic functions were assumed by ribozymes in the past,
then it should be possible to recreate such ribozymes in
the present. This prediction has been made implicitly or
explicitly by a number of authors, both prior to, and
following, the actual discovery of catalytic RNA [2,7-12].
In accord with these predictions, in vitro selection has
been used to generate a wide variety of new ribozymes.
Two types of selection procedure have yielded catalysts.
Indirect selection is similar to the protocols originally
used to isolate catalytic antibodies. Prudent et al. [13]
selected RNA molecules that can bind to a bridged
biphenyl analogue of the transition state of an isomeriza-
tion reaction. The binding species were found to be
ribozymes able to catalyze the isomerization of substrates
chemically similar to the analogue. Direct selection
requires that active RNA molecules in a pool alter them-
selves during the selection process in such a way as to
allow their isolation. For example, Bartel and Szostak [14]
were able to select RNA molecules able to ligate a primer
sequence onto themselves from a random sequence pool.
Conversely, Pan and Uhlenbeck [15] selected ribozymes
that can use a lead cofactor to cleave themselves.
Lorsch and Szostak [16] forged a link between cofactor-
binding and catalysis by selecting ribo-kinases that can
phosphorylate themselves from a pool of RNA mol-
ecules in which a randomly varied sequence was placed
next to a previously selected binding site for ATP. The
RNA world
selection of binding sites for redox cofactors similarly
opens the way to the selection of ribo-dehydrogenases or
ribo-oxidases [17,18]. Finally, direct selection for a
ribozyme that can catalyze a reaction that did not involve
a phosphodiester bond rearrangement has recently been
carried out by Wilson and Szostak [19]. These authors
identified ribozymes that can biotinylate themselves, an
alkylation reaction akin to those carried out by methyl
transferases throughout metabolism.
If at least some ribozymes were cobbled together from
random sequences during early evolution, then new
functions might have been quickly acquired by muta-
tional walks. Again, in vitro selection experiments give
credence to this idea. In particular, Joyce and his co-
workers have evolved the group I self-splicing intron to
perform reactions other than self-splicing, including
DNA cleavage [20-22] and RNA cleavage with calcium
[23]. Indeed, ribozymes that have evolved to catalyze one
reaction might intrinsically harbor the catalytic machin-
ery to carry out heterologous reactions. The group I self-
splicing intron has been shown to catalyze ester (as
opposed to phosphodiester) hydrolysis [24], and the vari-
ant evolved to catalyze DNA cleavage has been shown
also to hydrolyze amide bonds [25].
Amino acid-RNA interactions and the origin of
the genetic code
In the genetic code, the assignment of triplet codons to
particular amino acids is clearly non-random, yet the
logic or formula that could account for the standard
arrangement found in most organisms remains unknown.
Although it is possible that the genetic code is a 'frozen
accident' [7], its basic importance has driven various
authors to devise schemes that posit rational linkages
between genetic information and amino-acid functional-
ity. Perhaps the most beguiling of these hypotheses is the
idea that genetic information does not just code for, but
somehow begets, amino-acid functionality [26,27]. For
example, it has been proposed that amino acids may have
directly associated with their cognate triplet codons or
anticodons [28,29].
Once again, support for this hypothesis may be found in
the minutiae of biochemistry. The group I self-splicing
intron can interact specifically with a guanosine cofactor.
Yarus has shown that arginine can competitively inhibit
guanosine-binding to group I self-splicing introns [30],
and notes that the primary sequence of the guanosine-
binding site contains several trinucleotide tracts that could
be read as arginine codons [31]. This was interpreted to
be evidence in favor of the 'direct association' hypothesis,
as opposed to being just a fortuitous cross-reaction based
on structural similarities between the aptly named guano-
sine and the guanidino group of arginine (Fig. 2).
The generality of the amino-acid association observed
with the group I self-splicing intron can be readily
REVIEW 1019
Fig. 2. Structural similarities between arginine and guanosine. The
two structures are drawn so that the guanidino head group of argi-
nine is in a similar orientation to related chemical moieties found
on guanosine. The similarity can be extended through the base
and into the sugar, where either the primary amine or carboxylate
of the amino acid might overlap with hydroxyls on ribose.
assessed using in vitro selection. RNA molecules that can
specifically bind to arginine - 'aptamers' - were itera-
tively sieved from a completely random sequence pool
[32]. Aptamer sequences were determined and arginine-
binding sites mapped. The results obtained appeared to
be in accord with those observed for the group I intron.
One of the predominant aptamers did contain arginine
codons that overlapped with the arginine-binding site.
The ability of arginine to associate with RNA molecules
motivated Yarus [33,34] to propose a model for the origin
of the genetic code (Fig. 3). The model proposes that
guanosine-binding sites, such as those found in the group I
intron, were originally selected for during the evolution of
self-repli&tion. The structural similarities between guano-
sine and arginine allowed this amino acid to bind near the
active site of a ribozyme. This ribozyme may have been
able to use arginine as a substrate, catalyzing its activation
and then the aminoacylation of the ribozyme by the acti-
vated arginine. The arginine codon or anticodon (or both)
would have been a key feature of the binding site of the
newly evolved tRNA synthetase-like ribozyme. Other
tRNA synthetase-like ribozymes would have evolved with
their own coded interactions with amino acids. As pro-
teins displaced the RNA world, the ribozymes might have
faded, but the code would have remained.
Aspects of this model are supported by results from in
vitro selection experiments. The aptamers selected to bind
arginine were found also to bind guanosine. Aptamers
selected to bind both arginine and guanosine were found
to contain some arginine codons [35]. An RNA
1020 Current Biology 1995, Vol 5 No 9
Fig. 3. Model for the evolution of the translation apparatus and the
origin of the genetic code. This figure summarizes many of the
speculations put forth by Yarus and his co-workers [30,31,33,34].
Primordial replicases may have contained binding sites for nucleo-
tides such as guanosine (G) or adenosine (A). These binding sites
may have fortuitously also been able to bind some amino acids,
and could have served as 'proto-codons'. The ability of the guano-
sine-binding site of the Tetrahymena intron also to bind arginine
(R)is typically used as an example. Adjacent binding of amino acid
and nucleotide cofactors (A) could have resulted in the formation
of activated amino acids, such as amino acid adenylates. Activated
amino acids could in turn have reacted with terminal (or internal)
hydroxyls of the RNAs that held them. If these compounds or
events imparted function or phenotype to a ribo-organism, the
stage would have been set for the invention of translation. The
tRNA molecules seen in modern organisms can thus be viewed as
molecular fossils of ancient amino-acid-binding sites. The amazing
consistency of modern tRNA structures stands in stark contrast to
the apparent sequence and structural plasticity of selected amino-
acid-binding sites, and is not addressed by this model.
molecule that can catalyze an aminoacyl-RNA-synthe-
tase-like reaction using activated amino acids was selected
from a random sequence RNA pool. This demonstrates
the plausibility of the suggestion that ribozymes with
such catalytic activity may have existed in the RNA
world [36]. Taken together, these results affirm the plausi-
bility of the 'direct association' hypothesis and models for
the subsequent evolution of the genetic code.
Results from a similar number of selection experiments,
however, contradict these hypotheses and models. While
it is conceivable that one or more of the guanosine/
arginine-binding motifs gave rise to a tRNA synthetase-
like ribozyme, there are also multiple other ways to bind
arginine that do not necessarily involve arginine codons
(or anticodons). For example, Famulok [37] has selected
arginine-binding motifs that are not obviously dependent
on arginine codons or anticodons. More importantly, the
amino-acid specificity of these aptamers can be altered (to
citrulline) by substitutions at three residues, yet these
residues are not part of a single arginine codon or anti-
codon or codon-anticodon interaction. Moreover, there
is no indication from selection experiments that targeted
arginine-rich motifs on proteins [38], or other amino
acids such as tryptophan [39] or valine [40], that there is
any particular correspondence between selected motifs
and cognate codons or anticodons. If anything, selection
experiments seem to indicate that there are a huge num-
ber of possible binding motifs for a given amino acid.
Given the large number of possibilities, it may be impossi-
ble to prove or disprove any particular model for the evo-
lution of the genetic code. If the evolutionary 'film' were
to be run again, an entirely different, but equally deter-
mined, genetic code might be equally likely to emerge.
Antibiotics and the evolution of translation
Antibiotics are known to bind to, and disrupt the func-
tion of, ribosomal RNA. Conventionally, it was believed
that such interactions are due more to the structure of the
antibiotic than to the structure of the RNA. In this view,
antibiotics are compounds that were carefully crafted for
microbiological warfare and enhanced the survival of the
species that produced them. Julian Davies [41,42], how-
ever, has postulated that ribosomal RNA may be predis-
posed to interact with antibiotic inhibitors because of
historical constraints. For example, if ribosomes were
once regulated by small organic molecules (so-called low-
molecular-weight effectors or LMEs) then the binding
sites for these molecules may have been conserved
through the course of evolution, waiting to be exploited
by the combinatorial chemistry programs of bacteria.
More recently, it has been shown that antibiotics can bind
to, and inhibit, other functional RNAs, such as the group
I self-splicing intron and the hammerhead ribozyme
[43,44]. These results have led Renee Schroeder and
co-workers [45] to speculate that the active centers of
functional RNAs may form binding pockets that have
similar shapes or functions. In turn, if the ribosome and
the group I intron have similar LME-binding sites, then
they may also share a common ancestry [45,46]. This
novel hypothesis suggests that antibiotics can be used as
structural and functional probes to indicate relatedness,
much as antibodies were used to examine homologous
protein epitopes before the advent of phylogenetic
sequence analysis. Measuring the relatedness between
molecules such as ribosomal RNA and the group I intron
with a structural probe might be the only way by which
their ancestry could be confirmed, as these functional
RNAs are unrelated in primary and secondary structure.
RNA world
RNA world
Fig. 4. Models for the evolution of aminoglycoside-binding sites.
Two contrasting models for the presence of aminoglycoside-
binding sites in modern RNAs are shown. (a) The 'LME hypothe-
sis' suggests that modern aminoglycoside-binding sites are
molecular fossils of a relatively limited number of binding sites
that occurred in ancient RNAs. The sites have been conserved
through time because of their association with critical RNA struc-
tures or functions. (b) The 'stochastic hypothesis' suggests that, as
aminoglycosides have probably evolved to bind RNA, it is not
surprising that aminoglycoside-binding sites can be found by
chance alone on functional RNAs. Data from in vitro selection
experiments confirm that there are a wide variety of unnatural
aminoglycoside-binding sites 48,49].
The conjectures of Davies and Schroeder are particularly
intriguing because they tie together a variety of disparate
facts about functional RNAs and could validate the lineal
descent of modern organisms from ancient sequences.
The hypotheses must, however, be weighed against the
null hypothesis - that antibiotic-binding sites on differ-
ent RNAs are the result of chance rather than history.
This alternative is consistent with the fact that high-
affinity antibiotic-binding sites can be found on RNAs
that are probably not descended from primordial
sequences, such as the Rev-binding element of human
immunodeficiency virus 1 [47].
In vitro selection can be used to assess the likelihood of
the historical/LME hypothesis by assessing the likelihood
of its converse: the null (or 'stochastic') hypothesis (Fig.
4). If it is relatively difficult to find antibiotic-binding
sequences in a random sequence pool, then those anti-
biotic-binding sites that are identified in nature may be
due to common descent. If it is relatively easy to isolate
antibiotic-binding sequences, then there is no reason to
assume that binding sites that are otherwise unrelated
share a common ancestor.
REVIEW 1021
REVIEW 1021
Several groups have now carried out in vitro selection
experiments that target aminoglycoside antibiotics. Wang
and Rando [48] and Lato et al. [49] both find that even
limited sieving of random sequence pools can produce a
multitude of aptamers with moderate affinity
(Kd < 10 tIM) and specificity for a given aminoglycoside.
As has been shown for other targets, more extensive
selection results in the isolation of a limited number of
high-affinity (Kd < 1 FM) binding species. Even the
moderate-affinity aptamers can bind aminoglycosides as
well as ribosomal RNA and the group I self-splicing
intron can, and they can similarly discriminate between
related aminoglycosides.
These results suggest that natural aminoglycoside-binding
sites are far from unique, but are likely to be a subset of a
diverse universe of functionally related RNAs. However,
this result does not in itself necessarily invalidate the LME
hypothesis. For example, if there were distinguishing
characteristics that allowed aminoglycoside-binding apta-
mers to be parsed into classes, such as common primary
or secondary structural signatures, then it might still be
possible to show that natural binding sites are somehow
distinct from the plethora of unnatural binding sites.
Recently, Schroeder and her co-workers [50] have identi-
fied RNA motifs that can bind to neomycin, and suggest
that these motifs may contain a common (but not unique)
secondary structural feature - a widened major groove.
Comparisons and conclusions
The three examples examined above are purposefully
ordered in terms of chronology, specificity and implausi-
bility. This correspondence is not accidental: overall, the
main conclusion that can be drawn from in vitro selection
experiments is that the total number of potential binding
motifs and catalysts is gargantuan. Therefore, while it is
safe to speculate on the acquisition of a particular func-
tion or event (such as ribo-kinases or cellularization), the
very plurality of sequence and function may make it
impossible to prove the origins of a particular sequence
(the genetic code) or structure (the antibiotic-binding
site of the ribosome).
If this is the case, then it is not merely the information
within the genetic code that is a frozen accident of
biology, but perhaps all remnants of the inferred RNA
world. Cofactors, transfer RNAs and the ribosome
might all have turned out significantly different than
the versions we are familiar with, and deterministic
attempts to reconcile their sequences or structures with
one another or with other molecular traits, such as
self-splicing introns, become all the more difficult.
Paradoxically, examination of the precise nature of our
origins may be thwarted by the same factors that origi-
nally led to the success of the RNA world.
Acknowledgements: This work was supported by a National Science
Foundation National Young Investigator Award (A.E.), a Scholar
1022 Current Biology 1995, Vol 5 No 9
Award from the American Foundation for AIDS Research (A.E.),
the Pew Scholar Award in the Biomedical Sciences (A.E.), the
Cottrell Scholar Award (A.E.), the Office of Naval Research grant
#N00014-93-1-0430 (A.E.), and the National Institutes of Health
grant #PHS R01GM48175 (A.E.).
References
1. Gilbert W: The RNA world. Nature 1986, 319:618.
2. Benner SA, Ellington AD, Tauer A: Modern metabolism as a
palimpsest of the RNA world. Proc Natl Acad Sci USA 1989, 86:
7054-7058.
3. Gold L, Tuerk C: Methods for Identifying Nucleic Acid Ligands.
United States Patent #5270163, 1993.
4. Kramer FR, Mills DR, Cole PE, Nishihara T, Spiegelman S: Evolution
in vitro: sequence and phenotype of a mutant RNA resistant to
ethidium bromide. JMol Biol 1974, 89:719-736.
5. Orgel LE: Selection in vitro. Proc R Soc Lnd [Biol] 1979, 205:
435-442.
6. Visser CM: Evolution of bio catalysis 2. Nicotinamide and/or flavin-
containing RNA molecules as possible pre-genetic-code replicating
oxido-reductases. J Mol Evol 1984, 14:301-306.
7. Crick FHC: The origin of the genetic code. J Mol Biol 1968, 38:
367-379.
8. Orgel LE: Evolution of the genetic apparatus. Mol Biol 1968, 38:
381-393.
9. White HB: Coenzymes as fossils of an earlier metabolic state. JMol
Evol 1976, 7:101-104.
10. Visser CM, Kellogg RM: Biotin. Its place in evolution. Mol Evol
1978, 11:171-178.
11. Joyce GF: RNA evolution and the origins of life. Nature 1989,
338:21 7-224.
12. Benner SA, Cohen MA, Gonnet GH, Berkowitz DB, Johnsson KP:
Reading the palimpsest: contemporary biochemical data and
the RNA world. In The RNA World. Edited by Gesteland RF, Atkins
IF. New York: Cold Spring Harbor Laboratory Press; 1993:27-70.
13. Prudent JR, Uno T, Schultz PG: Expanding the scope of RNA cata-
lysis. Science 1994, 264:1924-1927.
14. Bartel DP, Szostak JW: Isolation of new ribozymes from a large pool
of random sequences. Science 1993, 261:1411-1418.
15. Pan T, Uhlenbeck OC: In vitro selection of RNAs that undergo
2+
autolytic cleavage with Pb . Biochemistry 1992, 31:3887-3995.
16. Lorsch JW, Szostak JW: In vitro evolution of new ribozymes with
polynucleotide kinase activity. Nature 1994, 371:31-36.
17. Burgstaller P, Famulok M: Isolation of RNA aptamers for biological
cofactors by in vitro selection. Angew Chem Int Ed Engl 1994,
33:1084-1087.
18. Lauhon CT, Szostak JW: RNA aptamers that bind flavin and nicotin-
amide redox cofactors. J Am Chem Soc 1995, 117:1246-1257.
19. Wilson C, Szostak JW: In vitro evolution of a self-alkylating
ribozyme. Nature 1995, 374:777-782.
20. Robertson DL, Joyce, GF: Selection in vitro of an RNA enzyme that
specifically cleaves single-stranded DNA. Nature 1990, 344:
467-468.
21. Beaudry A, Joyce GF: Directed evolution of an RNA enzyme.
Science 1992, 257:635-641.
22. Tsang J, Joyce GF: Evolutionary optimization of the catalytic
properties of a DNA-cleaving ribozyme. Biochemistry 1994, 33:
5966-5973.
23. Lehman N, Joyce GF: Evolution in vitro of an RNA enzyme with
altered metal dependence. Nature 1993, 361:182-185.
24. Piccirilli JA, McConnell TS, Zaug AJ, Noller HF, Cech TR: Amino-
acyl esterase activity of the Tetrahymena ribozyme. Science 1992,
256:1420-1424.
25. Dai X, Mesmaekder AD, Joyce GF: Cleavage of an amide bond by a
ribozyme. Science 1995, 267:237-240.
26. Szathmary E: Coding coenzyme handles: a hypothesis for the origin
of the genetic code. Proc Natl Acad Sci USA 1993, 90:
9916-9920.
27. Giulio MD, Capobianco MR, Medugno M: On the optimization of
the physiochemical distances between amino acids in the evolution
of the genetic code. Theor Biol 1994, 168:43-51.
28. Woese CR, Dugre DH, Dugre SA, Kondo M, Saxinger WC: On the
fundamental nature and evolution of the genetic code. Cold Spring
Harbor Symp Quant Biol 1966, 31:723-736.
29. Shimizu M: Molecular basis for the genetic code. J Mol Evol 1982,
18:297-303.
30. Yarus M: A specific amino-acid binding site composed of RNA.
Science 1988, 240:1751-1758.
31. Yarus M, Christian E: Genetic code origins. Nature 1989, 342:
349-350.
32. Connell GJ, Illangasekare M, Yarus M: Three small ribooligonucleo-
tides with specific arginine sites. Biochemistry 1993, 32:
5497-5502.
33. Yarus M: An RNA-amino acid complex and the origin of the
genetic code. New Biol 1991, 3:183-189.
34. Yarus M: An RNA-amino acid affinity. In The RNA World. Edited by
Gesteland RF, Atkins JF. New York: Cold Spring Harbor Laboratory
Press; 1993:205-217.
35. Connell GJ, Yarus M: RNAs with dual specificity and dual RNAs
with similar specificity. Science 1994, 264:1137-1141.
36. Illangasekare M, Sanchez G, Nickles T, Yarus M: Aminoacyl-RNA
synthesis catalyzed by an RNA. Science 1994, 267:643-647.
37. Famulok M: Molecular recognition of amino acids by RNA-
aptamers: an L-citrulline binding RNA motif and its evolution into
an L-arginine binder. J Am Chem Soc 1994, 116:1698-1706.
38. Giver L, Bartel D, Zapp M, Pawul A, Green M, Ellington AD: Selec-
tive optimization of the Rev-binding element of HIV-1. Nucleic
Acids Res 1993, 21:5509-5516.
39. Famulok M, Szostak JW: Stereospecific recognition of tryptophan
agarose by in vitro selected RNA. Am Chem Soc 1992 114:
3990-3991.
40. Majerfeld I, Yarus M: An RNA pocket for an aliphatic hydrophobe.
Nature Struct Biol 1994, 1:287-292.
41. Davies J: What are antibiotics? Archaic functions for modern activi-
ties. Mol Microbiol 1990, 4:1227-1232.
42. Davies J, von Ahsen U, Schroeder R: Antibiotics and the RNA
world: a role for low-molecular-weight effectors in biochemical
evolution? In The RNA World. Edited by Gesteland RF, Atkins F.
New York: Cold Spring Harbor Laboratory Press; 1993:185-204.
43. von Ahsen U, Davies J, Schroeder R: Antibiotic inhibition of group I
ribozyme function. Nature 1991, 353:368-370.
44. Stage TK, Hertel KJ,Uhlenbeck OC: Inhibition of the hammerhead
ribozyme by neomycin. RNA 1995, 1:95-101.
45. Schroeder R, Streicher B, Wank H: Splice-site selection and de-
coding: are they related? Science 1993, 260:1443-1444.
46. Noller HF: Drugs and the RNA world. Nature 1991, 353:302-303.
47. Zapp ML, Stern S, Green MR: Small molecules that selectively block
RNA binding of HIV-1 Rev protein inhibit Rev function and viral
production. Cell 1993, 74:969-978.
48. Wang Y, Rando RR: Specific binding of aminoglycoside antibiotics
to RNA. Chem & Biol 1995, 2:281-290.
49. Lato SM, Boles AR, Ellington AD: In vitro selection of RNA lectins:
using combinatorial chemistry to interpret ribozyme evolution.
Chem & Biol 1995, 2:291-303.
50. Wallis MG, von Ahsen U, Schroeder R, Famulok: A novel RNA
motif for neomycin recognition. Chem & Biol 1995, 2:543-552.
Received: 14 July 1995.