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PMID: 11497428 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Ribosomal protein L5 helps anchor peptidyl-tRNA to the P-site in Saccharomyces cerevisiae.

RNA (New York, N.Y.) ·Vol. 7 ·No. 8 ·2001-08-00 ·Pages 1084-96

Meskauskas A, Dinman JD

Abstract

Our previous demonstration that mutants of 5S rRNA called mof9 can specifically alter efficiencies of programmed ribosomal frameshifting (PRF) suggested a role for this ubiquitous molecule in the maintenance of translational reading frame, though the repetitive nature of the 5S rDNA gene (>100 copies/cell) inhibited more detailed analyses. However, given the known interactions between 5S rRNA and ribosomal protein L5 (previously called L1 or YL3) encoded by an essential, single-copy gene, we monitored the effects of a series of well-defined rpl5 mutants on PRF and virus propagation. Consistent with the mof9 results, we find that the rpl5 mutants promoted increased frameshifting efficiencies in both the -1 and +1 directions, and conferred defects in the ability of cells to propagate two endogenous viruses. Biochemical analyses demonstrated that mutant ribosomes had decreased affinities for peptidyl-tRNA. Pharmacological studies showed that sparsomycin, a peptidyltransferase inhibitor that specifically increases the binding of peptidyl-tRNA with ribosomes, was antagonistic to the frameshifting defects of the most severe mutant, and the extent of sparsomycin resistance correlated with the severity of the frameshifting defects in all of the mutants. These results provide biochemical and physiological evidence that one function of L5 is to anchor peptidyl-tRNA to the P-site. A model is presented describing how decreased affinity of ribosomes for peptidyl-tRNA can affect both -1 and +1 frameshifting, and for the effects of sparsomycin.

MeSH Terms
Alleles Anisomycin/pharmacology Antibiotics, Antineoplastic/pharmacology Dose-Response Relationship, Drug Frameshift Mutation Mutation Peptidyl Transferases/antagonists & inhibitors Phenotype Plasmids/metabolism Protein Biosynthesis Protein Synthesis Inhibitors/pharmacology RNA, Transfer/metabolism Retroelements/genetics Ribosomal Proteins/chemistry Ribosomes/metabolism Saccharomyces cerevisiae/metabolism Sparsomycin/pharmacology Time Factors
Chemicals
Antibiotics, Antineoplastic Protein Synthesis Inhibitors Retroelements Ribosomal Proteins ribosomal protein L5 Anisomycin Sparsomycin RNA, Transfer Peptidyl Transferases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Meskauskas A
Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway 08854, USA.
Dinman J D
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1017/s1355838201001480
Published
2001-08-00
Pages
1084-96
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1307509
Subset
IM
Grants
NIGMS NIH HHS · R01 GM058859 · United States
NIGMS NIH HHS · R01 GM062143 · United States
NIGMS NIH HHS · R01 GM58859 · United States
NIGMS NIH HHS · R01 GM62143 · United States
External Links
PubMed source DOI: 10.1017/s1355838201001480 PMC Full Text View journal details
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