A modified group I intron can function as both a ribozyme and a 5' exon in a trans-exon ligation reaction

Tasiouka, Konstantina I.; Burke, John M.

doi:10.1016/0378-1119(94)90195-3

Cited by 3 publications

(1 citation statement)

References 25 publications

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“…The discovery of RNAs with true enzymatic activities (ribozymes) (1,2) has provoked interest in exploring the catalytic potential of RNA. Meanwhile, various chemical reactions have been shown to be catalysed by members of the group I and group II intron ribozymes with substrates as different as RNA, DNA and even aminoacylated derivatives of RNA (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13). These results have repeatedly been discussed in the light of the role of RNA in a presumed RNA world (10,12,(14)(15)(16)(17)(18)(19).…”

Section: Introductionmentioning

confidence: 99%

DNA polymerization catalysed by a group II intron RNA in vitro

Hetzer

Schweyen

Mueller

1997

Nucleic Acids Research

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The excised group II intron bI1 from Saccharomyces cerevisiae can act as a ribozyme catalysing various chemical reactions with different substrate RNAs in vitro . Recently, we have described an editing-like RNA polymerization reaction catalysed by the bI1 intron lariat that proceeds in the 3'-->5'direction. Here we show that the bI1 lariat RNA can also catalyse successive deoxyribonucleotide polymerization reactions on exogenous substrate molecules. The basic mechanism of the reaction involved interacting cycles between an alternative version of partial reverse splicing (lariat charging) and canonical forward splicing (lariat discharging by exon ligation). With an overall chain growth in the 3'-->5' direction, the 5' exon RNAs (IBS1dN) were elongated by successive insertion of deoxyribonucleotides derived from single deoxyribonucleotide substitutions (dA, dG, dC or dT). All four deoxyribonucleotides were used as substrates, although with different efficiencies. Our findings extend the catalytic repertoire of group II intron RNAs not only by a novel DNA polymerization activity, but also by a DNA-DNA ligation capacity, supporting the idea that ribozymes might have been part of the first primordial polymerization machinery for both RNA and DNA.

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Section: Introductionmentioning

confidence: 99%

DNA polymerization catalysed by a group II intron RNA in vitro

Hetzer

Schweyen

Mueller

1997

Nucleic Acids Research

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Group I introns in biotechnology: prospects of application of ribozymes and rare-cutting homing endonucleases

Johansen

Einvik

Elde

et al. 1997

Biotechnology Annual Review

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Novel system for analysis of group I 3' splice site reactions based on functionaltrans-interaction of the P1/P10 reaction helix with the ribozyme's catalytic core

Chowrira¹,

Berzal‐Herranz²,

Burke³

1995

Nucl Acids Res

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A group I intron from a bacterial tRNA precursor has been converted into an RNA enzyme that catalyzes the efficient polymerization of oligoribonucleotide analogs of tRNA exons using a reaction scheme consisting of multiple cycles of reverse and forward exon ligation reactions. Here, we present results showing that this system represents a novel and useful tool for the analysis of 3' splice site reactions of group I ribozymes. First, analysis of variant substrates containing base substitutions in group I secondary structure elements P1, P9.0 and P10 confirms that exon polymerization is dependent on these structures, and therefore constitutes an appropriate and relevant model system for studying the exon ligation step of splicing. Second, to probe interactions between the intron's catalytic core and the bases and backbone of the P1/P10 reaction helix, two successful strategies for separating the internal guide sequence from the intron core were devised. One such strategy uses a construct in which the reaction helix interacts functionally with the catalytic core using only tertiary contacts. Further stabilization of this interaction through the inclusion of a 7 bp intermolecular P2 helix generates increased reaction efficiency. Third, when provided with two reaction helices, the ribozyme synthesizes mixed polymers through a mechanism that involves sequential binding and release of the duplexes. Fourth, in these reactions, turnover of the external guide sequence requires unwinding and annealing of the P2 helix, suggesting that P2 unwinding may occur during group I splicing. These results provide novel experimental tools to probe the relatively poorly understood 3' splice site reactions of group I introns, and may be relevant to ribozyme-catalyzed assembly and recombination of oligomers in prebiotic scenarios.

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A modified group I intron can function as both a ribozyme and a 5' exon in a trans-exon ligation reaction

Cited by 3 publications

References 25 publications

DNA polymerization catalysed by a group II intron RNA in vitro

DNA polymerization catalysed by a group II intron RNA in vitro

Group I introns in biotechnology: prospects of application of ribozymes and rare-cutting homing endonucleases

Novel system for analysis of group I 3' splice site reactions based on functionaltrans-interaction of the P1/P10 reaction helix with the ribozyme's catalytic core

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