• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. Jul 1995; 69(7): 4381–4389.
PMCID: PMC189179

Host cell proteins binding to domain IV of the 5' noncoding region of poliovirus RNA.

Abstract

Translation of poliovirus RNA occurs by the binding of ribosomes to an internal segment of RNA sequence within the 5' untranslated region of the viral RNA. This region is predicted to consist of six domains (I to VI) that possess complex secondary and tertiary structures. Domain IV is a large region in which alterations in the sequence or structure markedly reduce translational efficiency. In this study, we employed RNA mobility shift assays to demonstrate that a protein(s) from uninfected HeLa cell extracts, as well as from neuroblastoma extracts, interacts with the domain IV structure. A mutation in domain IV caused reduced binding of HeLa cell proteins and reduced translation both in vitro and in vivo, suggesting that the binding of at least one of these proteins plays a role in the mechanism of viral translation. UV cross-linking indicated that a protein(s) with a size of approximately 40 kDa interacted directly with the RNA. Using streptavidin beads to capture biotinylated RNA bound to proteins, we were able to visualize a number of HeLa and neuroblastoma cell proteins that interact with domain IV. These proteins have molecular masses of approximately 39, approximately 40, and approximately 42 kDa.

Full Text

The Full Text of this article is available as a PDF (528K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Alexander L, Lu HH, Wimmer E. Polioviruses containing picornavirus type 1 and/or type 2 internal ribosomal entry site elements: genetic hybrids and the expression of a foreign gene. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1406–1410. [PMC free article] [PubMed]
  • Andino R, Rieckhof GE, Baltimore D. A functional ribonucleoprotein complex forms around the 5' end of poliovirus RNA. Cell. 1990 Oct 19;63(2):369–380. [PubMed]
  • Borman A, Howell MT, Patton JG, Jackson RJ. The involvement of a spliceosome component in internal initiation of human rhinovirus RNA translation. J Gen Virol. 1993 Sep;74(Pt 9):1775–1788. [PubMed]
  • Borman AM, Deliat FG, Kean KM. Sequences within the poliovirus internal ribosome entry segment control viral RNA synthesis. EMBO J. 1994 Jul 1;13(13):3149–3157. [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]
  • Dildine SL, Semler BL. The deletion of 41 proximal nucleotides reverts a poliovirus mutant containing a temperature-sensitive lesion in the 5' noncoding region of genomic RNA. J Virol. 1989 Feb;63(2):847–862. [PMC free article] [PubMed]
  • Dildine SL, Semler BL. Conservation of RNA-protein interactions among picornaviruses. J Virol. 1992 Jul;66(7):4364–4376. [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]
  • Ehrenfeld E, Semler BL. Anatomy of the poliovirus internal ribosome entry site. Curr Top Microbiol Immunol. 1995;203:65–83. [PubMed]
  • Evans DM, Dunn G, Minor PD, Schild GC, Cann AJ, Stanway G, Almond JW, Currey K, Maizel JV., Jr Increased neurovirulence associated with a single nucleotide change in a noncoding region of the Sabin type 3 poliovaccine genome. Nature. 1985 Apr 11;314(6011):548–550. [PubMed]
  • Frankel AD, Mattaj IW, Rio DC. RNA-protein interactions. Cell. 1991 Dec 20;67(6):1041–1046. [PubMed]
  • Gabrielsen OS, Huet J. Magnetic DNA affinity purification of yeast transcription factor. Methods Enzymol. 1993;218:508–525. [PubMed]
  • Gebhard JR, Ehrenfeld E. Specific interactions of HeLa cell proteins with proposed translation domains of the poliovirus 5' noncoding region. J Virol. 1992 May;66(5):3101–3109. [PMC free article] [PubMed]
  • Haller AA, Nguyen JH, Semler BL. Minimum internal ribosome entry site required for poliovirus infectivity. J Virol. 1993 Dec;67(12):7461–7471. [PMC free article] [PubMed]
  • Haller AA, Semler BL. Linker scanning mutagenesis of the internal ribosome entry site of poliovirus RNA. J Virol. 1992 Aug;66(8):5075–5086. [PMC free article] [PubMed]
  • Haller AA, Semler BL. Stem-loop structure synergy in binding cellular proteins to the 5' noncoding region of poliovirus RNA. Virology. 1995 Feb 1;206(2):923–934. [PubMed]
  • Hellen CU, Pestova TV, Litterst M, Wimmer E. The cellular polypeptide p57 (pyrimidine tract-binding protein) binds to multiple sites in the poliovirus 5' nontranslated region. J Virol. 1994 Feb;68(2):941–950. [PMC free article] [PubMed]
  • Hellen CU, Witherell GW, Schmid M, Shin SH, Pestova TV, Gil A, Wimmer E. A cytoplasmic 57-kDa protein that is required for translation of picornavirus RNA by internal ribosomal entry is identical to the nuclear pyrimidine tract-binding protein. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7642–7646. [PMC free article] [PubMed]
  • Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. [PubMed]
  • HOLLAND JJ, McLAREN LC. Improved method for staining cell monolayers for virus plaque counts. J Bacteriol. 1959 Oct;78:596–597. [PMC free article] [PubMed]
  • Horton RM, Hunt HD, Ho SN, Pullen JK, Pease LR. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989 Apr 15;77(1):61–68. [PubMed]
  • Jackson RJ, Hunt SL, Gibbs CL, Kaminski A. Internal initiation of translation of picornavirus RNAs. Mol Biol Rep. 1994 May;19(3):147–159. [PubMed]
  • Jang SK, Kräusslich HG, Nicklin MJ, Duke GM, Palmenberg AC, Wimmer E. A segment of the 5' nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation. J Virol. 1988 Aug;62(8):2636–2643. [PMC free article] [PubMed]
  • Jang SK, Wimmer E. Cap-independent translation of encephalomyocarditis virus RNA: structural elements of the internal ribosomal entry site and involvement of a cellular 57-kD RNA-binding protein. Genes Dev. 1990 Sep;4(9):1560–1572. [PubMed]
  • Johnson VH, Semler BL. Defined recombinants of poliovirus and coxsackievirus: sequence-specific deletions and functional substitutions in the 5'-noncoding regions of viral RNAs. Virology. 1988 Jan;162(1):47–57. [PubMed]
  • Konarska MM, Sharp PA. Electrophoretic separation of complexes involved in the splicing of precursors to mRNAs. Cell. 1986 Sep 12;46(6):845–855. [PubMed]
  • Kuge S, Nomoto A. Construction of viable deletion and insertion mutants of the Sabin strain of type 1 poliovirus: function of the 5' noncoding sequence in viral replication. J Virol. 1987 May;61(5):1478–1487. [PMC free article] [PubMed]
  • La Monica N, Meriam C, Racaniello VR. Mapping of sequences required for mouse neurovirulence of poliovirus type 2 Lansing. J Virol. 1986 Feb;57(2):515–525. [PMC free article] [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]
  • Le SY, Zuker M. Common structures of the 5' non-coding RNA in enteroviruses and rhinoviruses. Thermodynamical stability and statistical significance. J Mol Biol. 1990 Dec 5;216(3):729–741. [PubMed]
  • Macadam AJ, Pollard SR, Ferguson G, Dunn G, Skuce R, Almond JW, Minor PD. The 5' noncoding region of the type 2 poliovirus vaccine strain contains determinants of attenuation and temperature sensitivity. Virology. 1991 Apr;181(2):451–458. [PubMed]
  • Meerovitch K, Pelletier J, Sonenberg N. A cellular protein that binds to the 5'-noncoding region of poliovirus RNA: implications for internal translation initiation. Genes Dev. 1989 Jul;3(7):1026–1034. [PubMed]
  • Meerovitch K, Svitkin YV, Lee HS, Lejbkowicz F, Kenan DJ, Chan EK, Agol VI, Keene JD, Sonenberg N. La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate. J Virol. 1993 Jul;67(7):3798–3807. [PMC free article] [PubMed]
  • Molla A, Paul AV, Wimmer E. Cell-free, de novo synthesis of poliovirus. Science. 1991 Dec 13;254(5038):1647–1651. [PubMed]
  • Molla A, Paul AV, Wimmer E. In vitro synthesis of poliovirus. Dev Biol Stand. 1993;78:39–53. [PubMed]
  • Omata T, Kohara M, Kuge S, Komatsu T, Abe S, Semler BL, Kameda A, Itoh H, Arita M, Wimmer E, et al. Genetic analysis of the attenuation phenotype of poliovirus type 1. J Virol. 1986 May;58(2):348–358. [PMC free article] [PubMed]
  • Pelletier J, Sonenberg N. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature. 1988 Jul 28;334(6180):320–325. [PubMed]
  • Percy N, Belsham GJ, Brangwyn JK, Sullivan M, Stone DM, Almond JW. Intracellular modifications induced by poliovirus reduce the requirement for structural motifs in the 5' noncoding region of the genome involved in internal initiation of protein synthesis. J Virol. 1992 Mar;66(3):1695–1701. [PMC free article] [PubMed]
  • Phillips BA, Emmert A. Modulation of the expression of poliovirus proteins in reticulocyte lysates. Virology. 1986 Jan 30;148(2):255–267. [PubMed]
  • Pilipenko EV, Blinov VM, Romanova LI, Sinyakov AN, Maslova SV, Agol VI. Conserved structural domains in the 5'-untranslated region of picornaviral genomes: an analysis of the segment controlling translation and neurovirulence. Virology. 1989 Feb;168(2):201–209. [PubMed]
  • Pöyry T, Kinnunen L, Hovi T. Genetic variation in vivo and proposed functional domains of the 5' noncoding region of poliovirus RNA. J Virol. 1992 Sep;66(9):5313–5319. [PMC free article] [PubMed]
  • Rivera VM, Welsh JD, Maizel JV., Jr Comparative sequence analysis of the 5' noncoding region of the enteroviruses and rhinoviruses. Virology. 1988 Jul;165(1):42–50. [PubMed]
  • Rohll JB, Percy N, Ley R, Evans DJ, Almond JW, Barclay WS. The 5'-untranslated regions of picornavirus RNAs contain independent functional domains essential for RNA replication and translation. J Virol. 1994 Jul;68(7):4384–4391. [PMC free article] [PubMed]
  • Ruby SW, Goelz SE, Hostomsky Z, Abelson JN. Affinity chromatography with biotinylated RNAs. Methods Enzymol. 1990;181:97–121. [PubMed]
  • Simoes EA, Sarnow P. An RNA hairpin at the extreme 5' end of the poliovirus RNA genome modulates viral translation in human cells. J Virol. 1991 Feb;65(2):913–921. [PMC free article] [PubMed]
  • Skinner MA, Racaniello VR, Dunn G, Cooper J, Minor PD, Almond JW. New model for the secondary structure of the 5' non-coding RNA of poliovirus is supported by biochemical and genetic data that also show that RNA secondary structure is important in neurovirulence. J Mol Biol. 1989 May 20;207(2):379–392. [PubMed]
  • Sonenberg N. Poliovirus translation. Curr Top Microbiol Immunol. 1990;161:23–47. [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, Cammack N, Minor PD, Almond JW. Translation deficiency of the Sabin type 3 poliovirus genome: association with an attenuating mutation C472----U. Virology. 1990 Mar;175(1):103–109. [PubMed]
  • Svitkin YV, Meerovitch K, Lee HS, Dholakia JN, Kenan DJ, Agol VI, Sonenberg N. Internal translation initiation on poliovirus RNA: further characterization of La function in poliovirus translation in vitro. J Virol. 1994 Mar;68(3):1544–1550. [PMC free article] [PubMed]
  • Trono D, Andino R, Baltimore D. An RNA sequence of hundreds of nucleotides at the 5' end of poliovirus RNA is involved in allowing viral protein synthesis. J Virol. 1988 Jul;62(7):2291–2299. [PMC free article] [PubMed]
  • Trono D, Pelletier J, Sonenberg N, Baltimore D. Translation in mammalian cells of a gene linked to the poliovirus 5' noncoding region. Science. 1988 Jul 22;241(4864):445–448. [PubMed]
  • Vaheri A, Pagano JS. Infectious poliovirus RNA: a sensitive method of assay. Virology. 1965 Nov;27(3):434–436. [PubMed]
  • Wimmer E, Hellen CU, Cao X. Genetics of poliovirus. Annu Rev Genet. 1993;27:353–436. [PubMed]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

Links

  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...