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PMID: 11387229 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transfer RNA(Ala) recognizes transfer-messenger RNA with specificity; a functional complex prior to entering the ribosome?

The EMBO journal ·Vol. 20 ·No. 11 ·2001-06-01 ·Pages 2966-76

Gillet R, Felden B

Abstract

tmRNA (SsrA or 10Sa RNA) functions as both a transfer RNA and a messenger RNA, rescues stalled ribosomes and clears the cell of incomplete polypeptides. We report that native Escherichia coli tmRNA interacts specifically with native or synthetic E.coli tRNA alanine (tRNA(Ala)) in vitro, alanine being the first codon of the tmRNA internal open reading frame. Aminoacylatable RNA microhelices also bind tmRNA. Complex formation was monitored by gel retardation assays combined with structural probes. Nucleotides from the acceptor stem of tRNA(Ala) are essential for complex formation with tmRNA. tRNA(Ala) isoacceptors recognize tmRNA with different affinities, with an important contribution from tRNA(Ala) post-transcriptional modifications. The most abundant tRNA(Ala) isoacceptor in vivo binds tmRNA with the highest affinity. A complex between tRNA(Ala) and tmRNA might involve up to 140 tmRNA molecules out of 500 present per E.coli cell. Our data suggest that tmRNA interacts with the tRNA that decodes the resume codon prior to entering the ribosome. Biological implications of promoting specific complexes between tmRNA and aminoacylatable RNAs are discussed, with emphasis on primitive versions of the translation apparatus.

MeSH Terms
Base Sequence Binding Sites Escherichia coli/genetics,metabolism Kinetics Molecular Sequence Data Nucleic Acid Conformation RNA, Bacterial/chemistry,metabolism RNA, Messenger/chemistry,metabolism RNA, Transfer, Ala/chemistry,metabolism RNA, Transfer, Asp/metabolism RNA, Transfer, Gln/metabolism Ribosomes/metabolism Substrate Specificity
Chemicals
RNA, Bacterial RNA, Messenger RNA, Transfer, Ala RNA, Transfer, Asp RNA, Transfer, Gln tmRNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gillet R
Laboratoire de Biochimie Pharmaceutique, Faculté de Pharmacie, Université de Rennes I, UPRES Jeune Equipe 2311, IFR 97, 2 avenue du Pr Léon Bernard, 35043 Rennes, France.
Felden B
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.11.2966
Published
2001-06-01
Pages
2966-76
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125487
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.11.2966 PMC Full Text View journal details
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