The Mechanism of an Exceptional Case of Reinitiation After Translation of a Long ORF Reveals Why Such Events Do Not Generally Occur in Mammalian MRNA Translation

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2007 Dec 7

PMID 18056426

Citations 58

Authors

Tuija A A Poyry

Ann Kaminski

Emma J Connell

Christopher S Fraser

Richard J Jackson

Affiliations

Soon will be listed here.

Abstract

The subgenomic mRNA of feline caliciviruses is bicistronic with the two cistrons overlapping by four nucleotides, ..AUGA. The upstream cistron encodes a 75-kDa major capsid protein precursor (pre-VP1), and the downstream cistron a 10-kDa minor capsid protein. The kinetics of translation in reticulocyte lysates show that the downstream cistron is translated by a termination-reinitiation process, which is unusual in not requiring eIF4G or the eIF4F complex. Reinitiation requires the 3'-terminal 87 nucleotides (nt) of the pre-VP1 ORF, but no other viral sequences. The reinitiation site is selected by virtue of its proximity to this 87-nt element, and not its proximity to the pre-VP1 ORF stop codon, although this must be located not more than approximately 30 nt downstream from the restart codon. This 87-nt element was shown to bind 40S ribosomal subunits and initiation factor eIF3, and addition of supplementary eIF3 enhanced reinitiation efficiency. Mutants defective in reinitiation showed reduced affinity for eIF3 or defective 40S subunit binding (or both). These results suggest a mechanism in which some of the eIF3/40S complexes formed during disassembly of post-termination ribosomes bind to this 87-nt element in a position appropriate for reinitiation following acquisition of an eIF2/GTP/Met-tRNA i ternary complex.

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References

Adhin M, van Duin J . Scanning model for translational reinitiation in eubacteria. J Mol Biol. 1990; 213(4):811-8. DOI: 10.1016/S0022-2836(05)80265-7. View

Peske F, Rodnina M, Wintermeyer W . Sequence of steps in ribosome recycling as defined by kinetic analysis. Mol Cell. 2005; 18(4):403-12. DOI: 10.1016/j.molcel.2005.04.009. View

Kojima K, Matsumoto T, Fujiwara H . Eukaryotic translational coupling in UAAUG stop-start codons for the bicistronic RNA translation of the non-long terminal repeat retrotransposon SART1. Mol Cell Biol. 2005; 25(17):7675-86. PMC: 1190309. DOI: 10.1128/MCB.25.17.7675-7686.2005. View

Kozak M . A short leader sequence impairs the fidelity of initiation by eukaryotic ribosomes. Gene Expr. 1991; 1(2):111-5. PMC: 5952205. View

Sosnovtsev S, Belliot G, Chang K, Onwudiwe O, Green K . Feline calicivirus VP2 is essential for the production of infectious virions. J Virol. 2005; 79(7):4012-24. PMC: 1061574. DOI: 10.1128/JVI.79.7.4012-4024.2005. View

Ali I, McKendrick L, Morley S, Jackson R . Truncated initiation factor eIF4G lacking an eIF4E binding site can support capped mRNA translation. EMBO J. 2001; 20(15):4233-42. PMC: 149147. DOI: 10.1093/emboj/20.15.4233. View

Kozak M . Constraints on reinitiation of translation in mammals. Nucleic Acids Res. 2002; 29(24):5226-32. PMC: 97554. DOI: 10.1093/nar/29.24.5226. View

Reynolds J, Kaminski A, Kettinen H, Grace K, Clarke B, Carroll A . Unique features of internal initiation of hepatitis C virus RNA translation. EMBO J. 1995; 14(23):6010-20. PMC: 394721. View

Sarabhai A, Brenner S . A mutant which reinitiates the polypeptide chain after chain termination. J Mol Biol. 1967; 27(1):145-62. DOI: 10.1016/0022-2836(67)90357-9. View

10.

Hunt S, Jackson R . Polypyrimidine-tract binding protein (PTB) is necessary, but not sufficient, for efficient internal initiation of translation of human rhinovirus-2 RNA. RNA. 1999; 5(3):344-59. PMC: 1369764. DOI: 10.1017/s1355838299981414. View

11.

Pestova T, Hellen C . Translation elongation after assembly of ribosomes on the Cricket paralysis virus internal ribosomal entry site without initiation factors or initiator tRNA. Genes Dev. 2003; 17(2):181-6. PMC: 195975. DOI: 10.1101/gad.1040803. View

12.

Steege D . 5'-Terminal nucleotide sequence of Escherichia coli lactose repressor mRNA: features of translational initiation and reinitiation sites. Proc Natl Acad Sci U S A. 1977; 74(10):4163-7. PMC: 431898. DOI: 10.1073/pnas.74.10.4163. View

13.

Pisarev A, Hellen C, Pestova T . Recycling of eukaryotic posttermination ribosomal complexes. Cell. 2007; 131(2):286-99. PMC: 2651563. DOI: 10.1016/j.cell.2007.08.041. View

14.

Gould P, Easton A . Coupled translation of the respiratory syncytial virus M2 open reading frames requires upstream sequences. J Biol Chem. 2005; 280(23):21972-80. DOI: 10.1074/jbc.M502276200. View

15.

Sosnovtsev S, Green K . Identification and genomic mapping of the ORF3 and VPg proteins in feline calicivirus virions. Virology. 2000; 277(1):193-203. DOI: 10.1006/viro.2000.0579. View

16.

Lomakin I, Shirokikh N, Yusupov M, Hellen C, Pestova T . The fidelity of translation initiation: reciprocal activities of eIF1, IF3 and YciH. EMBO J. 2005; 25(1):196-210. PMC: 1356347. DOI: 10.1038/sj.emboj.7600904. View

17.

Herbert T, Brierley I, Brown T . Identification of a protein linked to the genomic and subgenomic mRNAs of feline calicivirus and its role in translation. J Gen Virol. 1997; 78 ( Pt 5):1033-40. DOI: 10.1099/0022-1317-78-5-1033. View

18.

Wirblich C, Thiel H, Meyers G . Genetic map of the calicivirus rabbit hemorrhagic disease virus as deduced from in vitro translation studies. J Virol. 1996; 70(11):7974-83. PMC: 190870. DOI: 10.1128/JVI.70.11.7974-7983.1996. View

19.

Herbert T, Brierley I, Brown T . Detection of the ORF3 polypeptide of feline calicivirus in infected cells and evidence for its expression from a single, functionally bicistronic, subgenomic mRNA. J Gen Virol. 1996; 77 ( Pt 1):123-7. DOI: 10.1099/0022-1317-77-1-123. View

20.

Horvath C, Williams M, Lamb R . Eukaryotic coupled translation of tandem cistrons: identification of the influenza B virus BM2 polypeptide. EMBO J. 1990; 9(8):2639-47. PMC: 552297. View