Replacement of RNA hairpins by in vitro selected tetranucleotides

Bernhard Dichtl; Tao Pan; Anthony B. Direnzo; Olke C. Uhlenbeck

doi:10.1093/nar/21.3.531

Replacement of RNA hairpins by in vitro selected tetranucleotides

Bernhard Dichtl, Tao Pan, Anthony B. Direnzo, Olke C. Uhlenbeck^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

An in vitro selection method based on the autolytic cleavage of yeast tRNA^Phe by Pb²⁺ was applied to obtain tRNA derivatives with the anticodon hairpin replaced by four single-stranded nucleotides. Based on the rates of the site-specific cleavage by Pb²⁺ and the presence of a specific UV-induced crosslink, certain tetranucieotide sequences allow proper folding of the rest of the tRNA molecule, wheras others do not. One such successful tetramer sequence was also used to replace the acceptor stem of yeast tRNA^Phe and the anticodon hairpin of E.coli tRNA^Phe without disrupting folding. These experiments suggest that certain tetramers may be able to replace structurally non essential hairpins in any RNA.

Original language	English (US)
Pages (from-to)	531-535
Number of pages	5
Journal	Nucleic acids research
Volume	21
Issue number	3
DOIs	https://doi.org/10.1093/nar/21.3.531
State	Published - Feb 11 1993

Funding

This work was supported by N1H grant GM 37552. B.D. was partially supported by Deutscher Akademischer Austauschdienst (DAAD). T.P. acknowledges a postdoctoral fellowship from Damon Runyon-Walter Winchell Cancer Research Fund (DRG-1103). We thank J.Doudna and M.Stubenrauch for their comments on the manuscript. We also thank the W.M.Keck Foundation for their generous support of RNA science on the Boulder campus.

ASJC Scopus subject areas

Genetics

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title = "Replacement of RNA hairpins by in vitro selected tetranucleotides",

abstract = "An in vitro selection method based on the autolytic cleavage of yeast tRNAPhe by Pb2+ was applied to obtain tRNA derivatives with the anticodon hairpin replaced by four single-stranded nucleotides. Based on the rates of the site-specific cleavage by Pb2+ and the presence of a specific UV-induced crosslink, certain tetranucieotide sequences allow proper folding of the rest of the tRNA molecule, wheras others do not. One such successful tetramer sequence was also used to replace the acceptor stem of yeast tRNAPhe and the anticodon hairpin of E.coli tRNAPhe without disrupting folding. These experiments suggest that certain tetramers may be able to replace structurally non essential hairpins in any RNA.",

author = "Bernhard Dichtl and Tao Pan and Direnzo, {Anthony B.} and Uhlenbeck, {Olke C.}",

note = "Funding Information: This work was supported by N1H grant GM 37552. B.D. was partially supported by Deutscher Akademischer Austauschdienst (DAAD). T.P. acknowledges a postdoctoral fellowship from Damon Runyon-Walter Winchell Cancer Research Fund (DRG-1103). We thank J.Doudna and M.Stubenrauch for their comments on the manuscript. We also thank the W.M.Keck Foundation for their generous support of RNA science on the Boulder campus.",

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AU - Direnzo, Anthony B.

AU - Uhlenbeck, Olke C.

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N2 - An in vitro selection method based on the autolytic cleavage of yeast tRNAPhe by Pb2+ was applied to obtain tRNA derivatives with the anticodon hairpin replaced by four single-stranded nucleotides. Based on the rates of the site-specific cleavage by Pb2+ and the presence of a specific UV-induced crosslink, certain tetranucieotide sequences allow proper folding of the rest of the tRNA molecule, wheras others do not. One such successful tetramer sequence was also used to replace the acceptor stem of yeast tRNAPhe and the anticodon hairpin of E.coli tRNAPhe without disrupting folding. These experiments suggest that certain tetramers may be able to replace structurally non essential hairpins in any RNA.

AB - An in vitro selection method based on the autolytic cleavage of yeast tRNAPhe by Pb2+ was applied to obtain tRNA derivatives with the anticodon hairpin replaced by four single-stranded nucleotides. Based on the rates of the site-specific cleavage by Pb2+ and the presence of a specific UV-induced crosslink, certain tetranucieotide sequences allow proper folding of the rest of the tRNA molecule, wheras others do not. One such successful tetramer sequence was also used to replace the acceptor stem of yeast tRNAPhe and the anticodon hairpin of E.coli tRNAPhe without disrupting folding. These experiments suggest that certain tetramers may be able to replace structurally non essential hairpins in any RNA.

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Replacement of RNA hairpins by in vitro selected tetranucleotides

Abstract

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