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PMID: 6351012 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

The role of non-coding DNA sequences in transcription and processing of a yeast tRNA.

Nucleic acids research ·Vol. 11 ·No. 17 ·1983-09-10 ·Pages 5969-88

Raymond GJ, Johnson JD

Abstract

We have tested the hypothesis that conserved sequences in the intervening sequence (IVS) and 5'-flanking region of a yeast tRNALeu3 gene serve some function. Genes with deletions of 8, 10, 13 and 20 bp in the IVS are all active as templates in vitro. Yeast extracts produce mature tRNALeu3 from delta 8, delta 10 and delta 13 genes. Xenopus extracts do not detectably ligate the 5' and 3' half-molecules resulting from IVS excision. Neither extract is able to excise the IVS from delta 20 precursors. Genes with introns enlarged by 10, 21 or 30 bp of DNA produce mature tRNA. Insertion of 103 bp results in reduced levels of transcription, little if any end maturation, and no detectable mature product. A conserved 15 bp sequence is present at the 5'-end of the tRNA sequence. Replacement of yeast DNA up to position -22 leaves the tRNA gene transcriptionally active. With replacement extended to -2 the gene is active in Xenopus extracts but nearly inert in yeast extracts. We conclude that tRNA transcription in yeast is insensitive to IVS structure but can be positively influenced by 5'-flanking sequence.

MeSH Terms
Animals Base Sequence DNA, Fungal/genetics Female Genes Nucleic Acid Conformation Oocytes/metabolism Plasmids RNA, Transfer, Amino Acyl/genetics Saccharomyces cerevisiae/genetics Templates, Genetic Transcription, Genetic Xenopus
Chemicals
DNA, Fungal RNA, Transfer, Amino Acyl
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Raymond G J
Johnson J D
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46 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.17.5969
Published
1983-09-10
Pages
5969-88
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326330
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/11.17.5969 PMC Full Text View journal details
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