The C-terminal amino acid sequence of nascent peptide is a major determinant of SsrA tagging at all three stop codons

Takafumi Sunohara, Tatsuhiko Abo, Toshifumi Inada, Hiroji Aiba

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Recent studies on endogenous SsrA-tagged proteins have revealed that the tagging could occur at a position corresponding to the normal termination codon. During the study of SsrA-mediated Lacl tagging (Abo et al., EMBO J, 2000 19:3762-3769), we found that a variant Lacl (LaclΔC1) lacking the last C-terminal amino acid residue is efficiently tagged in a stop codon-dependent manner. SsrA tagging of LaclΔC1 occurred efficiently without Lacl binding to the lac operators at any one of three stop codons. The C-terminal (R) LESG peptide of LaclΔC1 was shown to trigger the SsrA tagging of an unrelated protein (CRP) when fused to its C terminus. Mass spectrometry analysis of the purified fusion proteins revealed that SsrA tagging occurs at a position corresponding to the termination codon. The alteration of the amino acid sequence but not the nucleotide sequence of the C-terminal portion eliminated the tagging. We also showed that the tagging-provoking sequences cause an efficient translational readthrough at UGA but not UAA codons. In addition, we found that C-terminal dipeptides known to induce an efficient translation readthrough could cause an efficient tagging at stop codons. We conclude that the amino acid sequence of nascent polypeptide prior to stop codons is a major determinant for the SsrA tagging at all three stop codons.

Original languageEnglish
Pages (from-to)1416-1427
Number of pages12
JournalRNA
Volume8
Issue number11
DOIs
Publication statusPublished - Nov 1 2002
Externally publishedYes

Keywords

  • Nascent peptide
  • Stop codon
  • TmRNA
  • Trans-translation
  • Translation readthrough

ASJC Scopus subject areas

  • Molecular Biology

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