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

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

Takafumi Sunohara et al. RNA. 2002 Nov.

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 (Lacl deltaC1) lacking the last C-terminal amino acid residue is efficiently tagged in a stop codon-dependent manner. SsrA tagging of Lacl deltaC1 occurred efficiently without Lacl binding to the lac operators at any one of three stop codons. The C-terminal (R)LESG peptide of Lacl deltaC1 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.

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References

1. Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5 - PubMed
1. EMBO J. 1999 Aug 16;18(16):4579-89 - PubMed
1. EMBO J. 2000 Jul 17;19(14):3762-9 - PubMed
1. Science. 2000 Aug 11;289(5481):905-20 - PubMed
1. Science. 2000 Aug 11;289(5481):920-30 - PubMed

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