Site-specific release of nascent chains from ribosomes at a sense codon

被引:126
作者
Doronina, Victoria A. [1 ,2 ]
Wu, Cheng [3 ]
de Felipe, Pablo [4 ]
Sachs, Matthew S. [3 ,5 ]
Ryan, Martin D. [4 ]
Brown, Jeremy D. [1 ,2 ]
机构
[1] Univ Newcastle Upon Tyne, Sch Med, Inst Cell & Mol Biosci, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
[2] Univ Newcastle Upon Tyne, Sch Med, RNA Biol Grp, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
[3] Oregon Hlth & Sci Univ, Dept Environm & Biomol Syst, Beaverton, OR 97006 USA
[4] Univ St Andrews, Ctr Biomol Sci, Sch Biol, St Andrews KY16 9ST, Fife, Scotland
[5] Oregon Hlth & Sci Univ, Dept Mol Microbiol & Immunol, Portland, OR 97201 USA
基金
英国生物技术与生命科学研究理事会; 英国医学研究理事会;
关键词
D O I
10.1128/MCB.00421-08
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
2A oligopeptides are autonomous elements containing a D(V/I)EXNPGP motif at the C terminus. Protein synthesis from an open reading frame containing an internal 2A coding sequence yields two separate polypeptides, corresponding to sequences up to and including 2A and those downstream. We show that the 2A reaction occurs in the ribosomal peptidyltransferase center. Ribosomes pause at the end of the 2A coding sequence, over the glycine and proline codons, and the nascent chain up to and including this glycine is released. Translation-terminating release factors eRF1 and eRF3 play key roles in the reaction. On the depletion of eRF1, a greater proportion of ribosomes extend through the 2A coding sequence, yielding the full-length protein. In contrast, impaired eRF3 GTPase activity leads to many ribosomes failing to translate beyond 2A. Further, high-level expression of a 2A peptide-containing protein inhibits the growth of cells compromised for release factor activity and leads to errors in stop codon recognition. We propose that the nascent 2A peptide interacts with ribosomes to drive a highly unusual and specific "termination" reaction, despite the presence of a proline codon in the A site. After this, the majority of ribosomes continue translation, generating the separate downstream product.
引用
收藏
页码:4227 / 4239
页数:13
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