A single base change in the intron of a serine tRNA affects the rate of RNase P cleavage in vitro and suppressor activity in vivo in Saccharomyces cerevisiae

Ian M. Willis, D. Frendewey, M. Nichols, A. Hottinger-Werlen, J. Schaack, D. Söll

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Differences in the processing of dimeric tRNA(Ser)-tRNA(Met) precursors derived from the Schizosaccharomyces pombe sup9 wild-type and opal suppressor genes can be attributed to conformational alterations in the tRNA(Ser) anticodon/intron domain. A comparison of the patterns obtained upon transcription of the sup9+ (wild-type) and sup9-e (opal suppressor) genes in a coupled transcription/processing extract from Saccharomyces cerevisiae reveals that the latter exhibits a greatly reduced efficiency of 5'-end maturation and is susceptible to specific endonucleolytic cleavage(s) within the intron. Free energy calculations indicate that these effects coincide with a destabilization of the wild-type anticodon/intron stem and suggest that the predominant sup9-e conformer lacks secondary structure in this region. Evidence in support of this hypothesis was obtained by (i) analyzing the processing of sup9+ and sup9-e precursors carrying the intron base substitution, G37:10, which destroys and restores, respectively, the base-pairing potential of the proposed secondary structure and (ii) comparing the strength and temperature sensitivity of sup9-e and sup9-e G37:10 suppression in vivo in S. cerevisiae. The data indicate that the anticodon/intron structure of tRNA precursors can influence the rate of RNase P cleavage in vitro and affect tRNA expression in vivo.

Original languageEnglish (US)
Pages (from-to)5878-5885
Number of pages8
JournalJournal of Biological Chemistry
Volume261
Issue number13
StatePublished - 1986
Externally publishedYes

Fingerprint

Ribonuclease P
Transfer RNA
Yeast
Introns
Serine
Saccharomyces cerevisiae
Anticodon
RNA, Transfer, Ser
Suppressor Genes
RNA Precursors
Transcription
Processing
Genes
RNA, Transfer, Met
Schizosaccharomyces
Base Pairing
Free energy
Substitution reactions
In Vitro Techniques
Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

A single base change in the intron of a serine tRNA affects the rate of RNase P cleavage in vitro and suppressor activity in vivo in Saccharomyces cerevisiae. / Willis, Ian M.; Frendewey, D.; Nichols, M.; Hottinger-Werlen, A.; Schaack, J.; Söll, D.

In: Journal of Biological Chemistry, Vol. 261, No. 13, 1986, p. 5878-5885.

Research output: Contribution to journalArticle

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AU - Willis, Ian M.

AU - Frendewey, D.

AU - Nichols, M.

AU - Hottinger-Werlen, A.

AU - Schaack, J.

AU - Söll, D.

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