• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of rnaThe RNA SocietyeTOC AlertsSubscriptionsJournal HomeCSHL PressRNA
RNA. May 1998; 4(5): 520–529.
PMCID: PMC1369636

Recognition of picornavirus internal ribosome entry sites within cells; influence of cellular and viral proteins.

Abstract

The ability of different picornavirus internal ribosome entry site (IRES) elements to direct initiation of protein synthesis has been assayed in different cell lines in the presence and absence of viral proteases that inhibit cap-dependent protein synthesis. Reporter plasmids that express dicistronic mRNAs, containing different IRES elements, with the general structure CAT/IRES/LUC, have been assayed. In each plasmid, the CAT sequence encodes chloramphenicol acetyl transferase and the LUC sequence encodes luciferase. The poliovirus (PV) 2A protease and the foot-and-mouth disease virus (FMDV) Lb protease induce the cleavage of the translation initiation factor elF4G and hence inhibit the activity of the cap-binding complex, elF4F. In human osteosarcoma (HTK-143) cells, each of the various IRES elements functioned efficiently. In these cells, the co-expression of the viral proteases severely inhibited the expression of CAT, but the proteases had little effect on the activities of the various IRES elements. In contrast, in baby hamster kidney (BHK) cells, the efficiencies of the different IRES elements varied significantly, whereas, in normal rat kidney (NRK) cells, each of the IRES elements was relatively inefficient. In both BHK and NRK cells, the activities of those IRES elements that functioned inefficiently were strongly stimulated by the co-expression of the PV 2A or FMDV Lb proteases. This stimulation was independent of the loss of cap-dependent protein synthesis and was not achieved by the co-expression of the C-terminal fragment of elF4G. The results suggest that the PV 2A and FMDV Lb proteases induce the cleavage of another cellular protein, in addition to elF4G, which influences IRES function.

Full Text

The Full Text of this article is available as a PDF (610K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Agol VI, Drozdov SG, Ivannikova TA, Kolesnikova MS, Korolev MB, Tolskaya EA. Restricted growth of attenuated poliovirus strains in cultured cells of a human neuroblastoma. J Virol. 1989 Sep;63(9):4034–4038. [PMC free article] [PubMed]
  • Belsham GJ, Brangwyn JK. A region of the 5' noncoding region of foot-and-mouth disease virus RNA directs efficient internal initiation of protein synthesis within cells: involvement with the role of L protease in translational control. J Virol. 1990 Nov;64(11):5389–5395. [PMC free article] [PubMed]
  • Belsham GJ, Sonenberg N. RNA-protein interactions in regulation of picornavirus RNA translation. Microbiol Rev. 1996 Sep;60(3):499–511. [PMC free article] [PubMed]
  • Borman AM, Kirchweger R, Ziegler E, Rhoads RE, Skern T, Kean KM. elF4G and its proteolytic cleavage products: effect on initiation of protein synthesis from capped, uncapped, and IRES-containing mRNAs. RNA. 1997 Feb;3(2):186–196. [PMC free article] [PubMed]
  • Borman AM, Le Mercier P, Girard M, Kean KM. Comparison of picornaviral IRES-driven internal initiation of translation in cultured cells of different origins. Nucleic Acids Res. 1997 Mar 1;25(5):925–932. [PMC free article] [PubMed]
  • Brown BA, Ehrenfeld E. Translation of poliovirus RNA in vitro: changes in cleavage pattern and initiation sites by ribosomal salt wash. Virology. 1979 Sep;97(2):396–405. [PubMed]
  • Devaney MA, Vakharia VN, Lloyd RE, Ehrenfeld E, Grubman MJ. Leader protein of foot-and-mouth disease virus is required for cleavage of the p220 component of the cap-binding protein complex. J Virol. 1988 Nov;62(11):4407–4409. [PMC free article] [PubMed]
  • Dorner AJ, Semler BL, Jackson RJ, Hanecak R, Duprey E, Wimmer E. In vitro translation of poliovirus RNA: utilization of internal initiation sites in reticulocyte lysate. J Virol. 1984 May;50(2):507–514. [PMC free article] [PubMed]
  • Drew J, Belsham GJ. trans complementation by RNA of defective foot-and-mouth disease virus internal ribosome entry site elements. J Virol. 1994 Feb;68(2):697–703. [PMC free article] [PubMed]
  • Fuerst TR, Niles EG, Studier FW, Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122–8126. [PMC free article] [PubMed]
  • Glass MJ, Summers DF. Identification of a trans-acting activity from liver that stimulates hepatitis A virus translation in vitro. Virology. 1993 Apr;193(2):1047–1050. [PubMed]
  • Haghighat A, Mader S, Pause A, Sonenberg N. Repression of cap-dependent translation by 4E-binding protein 1: competition with p220 for binding to eukaryotic initiation factor-4E. EMBO J. 1995 Nov 15;14(22):5701–5709. [PMC free article] [PubMed]
  • Hambidge SJ, Sarnow P. Translational enhancement of the poliovirus 5' noncoding region mediated by virus-encoded polypeptide 2A. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10272–10276. [PMC free article] [PubMed]
  • Jackson RJ, Kaminski A. Internal initiation of translation in eukaryotes: the picornavirus paradigm and beyond. RNA. 1995 Dec;1(10):985–1000. [PMC free article] [PubMed]
  • Kaminski A, Howell MT, Jackson RJ. Initiation of encephalomyocarditis virus RNA translation: the authentic initiation site is not selected by a scanning mechanism. EMBO J. 1990 Nov;9(11):3753–3759. [PMC free article] [PubMed]
  • Kirchweger R, Ziegler E, Lamphear BJ, Waters D, Liebig HD, Sommergruber W, Sobrino F, Hohenadl C, Blaas D, Rhoads RE, et al. Foot-and-mouth disease virus leader proteinase: purification of the Lb form and determination of its cleavage site on eIF-4 gamma. J Virol. 1994 Sep;68(9):5677–5684. [PMC free article] [PubMed]
  • Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. [PMC free article] [PubMed]
  • Kräusslich HG, Nicklin MJ, Toyoda H, Etchison D, Wimmer E. Poliovirus proteinase 2A induces cleavage of eucaryotic initiation factor 4F polypeptide p220. J Virol. 1987 Sep;61(9):2711–2718. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • La Monica N, Racaniello VR. Differences in replication of attenuated and neurovirulent polioviruses in human neuroblastoma cell line SH-SY5Y. J Virol. 1989 May;63(5):2357–2360. [PMC free article] [PubMed]
  • Lamphear BJ, Kirchweger R, Skern T, Rhoads RE. Mapping of functional domains in eukaryotic protein synthesis initiation factor 4G (eIF4G) with picornaviral proteases. Implications for cap-dependent and cap-independent translational initiation. J Biol Chem. 1995 Sep 15;270(37):21975–21983. [PubMed]
  • Lamphear BJ, Yan R, Yang F, Waters D, Liebig HD, Klump H, Kuechler E, Skern T, Rhoads RE. Mapping the cleavage site in protein synthesis initiation factor eIF-4 gamma of the 2A proteases from human Coxsackievirus and rhinovirus. J Biol Chem. 1993 Sep 15;268(26):19200–19203. [PubMed]
  • Macadam AJ, Ferguson G, Fleming T, Stone DM, Almond JW, Minor PD. Role for poliovirus protease 2A in cap independent translation. EMBO J. 1994 Feb 15;13(4):924–927. [PMC free article] [PubMed]
  • Mader S, Lee H, Pause A, Sonenberg N. The translation initiation factor eIF-4E binds to a common motif shared by the translation factor eIF-4 gamma and the translational repressors 4E-binding proteins. Mol Cell Biol. 1995 Sep;15(9):4990–4997. [PMC free article] [PubMed]
  • Medina M, Domingo E, Brangwyn JK, Belsham GJ. The two species of the foot-and-mouth disease virus leader protein, expressed individually, exhibit the same activities. Virology. 1993 May;194(1):355–359. [PubMed]
  • Ohlmann T, Pain VM, Wood W, Rau M, Morley SJ. The proteolytic cleavage of eukaryotic initiation factor (eIF) 4G is prevented by eIF4E binding protein (PHAS-I; 4E-BP1) in the reticulocyte lysate. EMBO J. 1997 Feb 17;16(4):844–855. [PMC free article] [PubMed]
  • Ohlmann T, Rau M, Pain VM, Morley SJ. The C-terminal domain of eukaryotic protein synthesis initiation factor (eIF) 4G is sufficient to support cap-independent translation in the absence of eIF4E. EMBO J. 1996 Mar 15;15(6):1371–1382. [PMC free article] [PubMed]
  • Pause A, Belsham GJ, Gingras AC, Donzé O, Lin TA, Lawrence JC, Jr, Sonenberg N. Insulin-dependent stimulation of protein synthesis by phosphorylation of a regulator of 5'-cap function. Nature. 1994 Oct 27;371(6500):762–767. [PubMed]
  • Pestova TV, Shatsky IN, Hellen CU. Functional dissection of eukaryotic initiation factor 4F: the 4A subunit and the central domain of the 4G subunit are sufficient to mediate internal entry of 43S preinitiation complexes. Mol Cell Biol. 1996 Dec;16(12):6870–6878. [PMC free article] [PubMed]
  • Pollard SR, Dunn G, Cammack N, Minor PD, Almond JW. Nucleotide sequence of a neurovirulent variant of the type 2 oral poliovirus vaccine. J Virol. 1989 Nov;63(11):4949–4951. [PMC free article] [PubMed]
  • Roberts LO, Belsham GJ. Complementation of defective picornavirus internal ribosome entry site (IRES) elements by the coexpression of fragments of the IRES. Virology. 1997 Jan 6;227(1):53–62. [PubMed]
  • Stone DM, Almond JW, Brangwyn JK, Belsham GJ. trans complementation of cap-independent translation directed by poliovirus 5' noncoding region deletion mutants: evidence for RNA-RNA interactions. J Virol. 1993 Oct;67(10):6215–6223. [PMC free article] [PubMed]
  • Svitkin YV, Maslova SV, Agol VI. The genomes of attenuated and virulent poliovirus strains differ in their in vitro translation efficiencies. Virology. 1985 Dec;147(2):243–252. [PubMed]
  • Svitkin YV, Pestova TV, Maslova SV, Agol VI. Point mutations modify the response of poliovirus RNA to a translation initiation factor: a comparison of neurovirulent and attenuated strains. Virology. 1988 Oct;166(2):394–404. [PubMed]
  • Van Der Velden A, Kaminski A, Jackson RJ, Belsham GJ. Defective point mutants of the encephalomyocarditis virus internal ribosome entry site can be complemented in trans. Virology. 1995 Dec 1;214(1):82–90. [PubMed]
  • Yamanaka S, Poksay KS, Arnold KS, Innerarity TL. A novel translational repressor mRNA is edited extensively in livers containing tumors caused by the transgene expression of the apoB mRNA-editing enzyme. Genes Dev. 1997 Feb 1;11(3):321–333. [PubMed]
  • Ziegler E, Borman AM, Deliat FG, Liebig HD, Jugovic D, Kean KM, Skern T, Kuechler E. Picornavirus 2A proteinase-mediated stimulation of internal initiation of translation is dependent on enzymatic activity and the cleavage products of cellular proteins. Virology. 1995 Nov 10;213(2):549–557. [PubMed]
  • Ziegler E, Borman AM, Kirchweger R, Skern T, Kean KM. Foot-and-mouth disease virus Lb proteinase can stimulate rhinovirus and enterovirus IRES-driven translation and cleave several proteins of cellular and viral origin. J Virol. 1995 Jun;69(6):3465–3474. [PMC free article] [PubMed]

Articles from RNA are provided here courtesy of The RNA Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...