Cap-Poly(A) synergy in mammalian cell-free extracts. Investigation of the requirements for poly(A)-mediated stimulation of translation initiation - PubMed

Affiliations

• PMID: 10922367
• DOI: 10.1074/jbc.M004304200

Free article

Cap-Poly(A) synergy in mammalian cell-free extracts. Investigation of the requirements for poly(A)-mediated stimulation of translation initiation

Y M Michel et al. J Biol Chem. 2000.

Free article

Abstract

The 5' cap and 3' poly(A) tail of eukaryotic mRNAs cooperate to stimulate synergistically translation initiation in vivo, a phenomenon observed to date in vitro only in translation systems containing endogenous competitor mRNAs. Here we describe nuclease-treated rabbit reticulocyte lysates and HeLa cell cytoplasmic extracts that reproduce cap-poly(A) synergy in the absence of such competitor RNAs. Extracts were rendered poly(A)-dependent by ultracentrifugation to partially deplete them of ribosomes and associated initiation factors. Under optimal conditions, values for synergy in reticulocyte lysates approached 10-fold. By using this system, we investigated the molecular mechanism of poly(A) stimulation of translation. Maximal cap-poly(A) cooperativity required the integrity of the eukaryotic initiation factor 4G-poly(A)-binding protein (eIF4G-PABP) interaction, suggesting that synergy results from mRNA circularization. In addition, polyadenylation stimulated uncapped cellular mRNA translation and that driven by the encephalomyocarditis virus internal ribosome entry segment (IRES). These effects of poly(A) were also sensitive to disruption of the eIF4G-PABP interaction, suggesting that 5'-3' end cross-talk is functionally conserved between classical mRNAs and an IRES-containing mRNA. Finally, we demonstrate that a rotaviral non-structural protein that evicts PABP from eIF4G is capable of provoking the shut-off of host cell translation seen during rotavirus infection.

Similar articles

• Biochemical characterisation of cap-poly(A) synergy in rabbit reticulocyte lysates: the eIF4G-PABP interaction increases the functional affinity of eIF4E for the capped mRNA 5'-end.

  Borman AM, Michel YM, Kean KM. Borman AM, et al. Nucleic Acids Res. 2000 Nov 1;28(21):4068-75. doi: 10.1093/nar/28.21.4068. Nucleic Acids Res. 2000. PMID: 11058101 Free PMC article.

• Back to basics: the untreated rabbit reticulocyte lysate as a competitive system to recapitulate cap/poly(A) synergy and the selective advantage of IRES-driven translation.

  Soto Rifo R, Ricci EP, Décimo D, Moncorgé O, Ohlmann T. Soto Rifo R, et al. Nucleic Acids Res. 2007;35(18):e121. doi: 10.1093/nar/gkm682. Epub 2007 Sep 18. Nucleic Acids Res. 2007. PMID: 17881372 Free PMC article.

• [Translational control by the poly(A) binding protein: a check for mRNA integrity].

  Svitkin YV, Sonenberg N. Svitkin YV, et al. Mol Biol (Mosk). 2006 Jul-Aug;40(4):684-93. Mol Biol (Mosk). 2006. PMID: 16913227 Review. Russian.

Cited by

• Preferential translation of vesicular stomatitis virus mRNAs is conferred by transcription from the viral genome.

  Whitlow ZW, Connor JH, Lyles DS. Whitlow ZW, et al. J Virol. 2006 Dec;80(23):11733-42. doi: 10.1128/JVI.00971-06. Epub 2006 Sep 27. J Virol. 2006. PMID: 17005665 Free PMC article.

• Yeast transcripts cleaved by an internal ribozyme provide new insight into the role of the cap and poly(A) tail in translation and mRNA decay.

  Meaux S, Van Hoof A. Meaux S, et al. RNA. 2006 Jul;12(7):1323-37. doi: 10.1261/rna.46306. Epub 2006 May 19. RNA. 2006. PMID: 16714281 Free PMC article.

• Detailed analysis of the requirements of hepatitis A virus internal ribosome entry segment for the eukaryotic initiation factor complex eIF4F.

  Borman AM, Michel YM, Kean KM. Borman AM, et al. J Virol. 2001 Sep;75(17):7864-71. doi: 10.1128/jvi.75.17.7864-7871.2001. J Virol. 2001. PMID: 11483730 Free PMC article.

• Competitive translation efficiency at the picornavirus type 1 internal ribosome entry site facilitated by viral cis and trans factors.

  Dobrikova EY, Grisham RN, Kaiser C, Lin J, Gromeier M. Dobrikova EY, et al. J Virol. 2006 Apr;80(7):3310-21. doi: 10.1128/JVI.80.7.3310-3321.2006. J Virol. 2006. PMID: 16537598 Free PMC article.

• General RNA-binding proteins have a function in poly(A)-binding protein-dependent translation.

  Svitkin YV, Evdokimova VM, Brasey A, Pestova TV, Fantus D, Yanagiya A, Imataka H, Skabkin MA, Ovchinnikov LP, Merrick WC, Sonenberg N. Svitkin YV, et al. EMBO J. 2009 Jan 7;28(1):58-68. doi: 10.1038/emboj.2008.259. Epub 2008 Dec 11. EMBO J. 2009. PMID: 19078965 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Elsevier Science

• Other Literature Sources

  • The Lens - Patent Citations Database

• Miscellaneous

  • NCI CPTAC Assay Portal