Contributions of Multiple Basic Amino Acids in the C-terminal Region of Yeast Ribosomal Protein L1 to 5 S rRNA Binding and 60 S Ribosome Stability

Lee Chuan C. Yeh, John C. Lee

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Previous studies suggest that the C-terminal region of ribosomal protein L1 fromSaccharomyces cerevisiaeis important for its interaction with the 5 S rRNA molecule. Within this region are several highly conserved basic amino acids including Lys276, Lys279, Lys289, Arg282, Arg285. To examine potential contributions of these amino acids to RNA-protein interaction and ribosomal assembly, effects of substitutions of these residues by methionine either individually or in combinations were examined. A methionine substitution of any one of the lysine residues did not significantly affect RNA bindingin vitro. The mutant RNPs were as stable as the wild-type RNP. Yeast transformants expressing these mutant proteins grew at the same rate as the wild-type. However, mutant proteins containing substitutions of any two of these basic amino acids bound RNA weakly. The resultant RNPs were significantly less stable than the wild-type. Whereas cells expressing mutant L1 with a single substitution at 289 was not lethal, cells expressing mutant L1 with any double substitutions involving Lys289 as one of the substituted amino acids were lethal. These data suggest that Lys289 plays a key role in the binding of ribosomal protein L1 to 5 S rRNA. The other basic residues, particularly Arg282, and Arg285, in this region also contribute to RNA binding. These residues are predicted to locate on the same side of an α helix. We would like to propose a structural model for the yeast RNP that involves multiple contact sites located on one side of the helix in the C terminus of the protein and the 5 S rRNA. These basic amino acids also participate, directly or indirectly, in the interaction of the RNP complex with other components of the 60 S ribosomal subunit.

Original languageEnglish (US)
Pages (from-to)295-307
Number of pages13
JournalJournal of Molecular Biology
Volume246
Issue number2
DOIs
StatePublished - Feb 17 1995

Keywords

  • 5 S rRNA
  • RNA-protein interaction
  • Ribosomal protein
  • Ribosome
  • Yeast

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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