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. 1997 Jan;71(1):578–585. doi: 10.1128/jvi.71.1.578-585.1997

Processing of a cellular polypeptide by 3CD proteinase is required for poliovirus ribonucleoprotein complex formation.

H H Roehl 1, T B Parsley 1, T V Ho 1, B L Semler 1
PMCID: PMC191087  PMID: 8985386

Abstract

Poliovirus interactions with host cells were investigated by studying the formation of ribonucleoprotein complexes at the 3' end of poliovirus negative-strand RNA which are presumed to be involved in viral RNA synthesis. It was previously shown that two host cell proteins with molecular masses of 36 and 38 kDa bind to the 3' end of viral negative-strand RNA at approximately 3 to 4 h after infection. We tested the hypothesis that preexisting cellular proteins are modified during the course of infection and are subsequently recruited to play a role in viral replication. It was demonstrated that the 38-kDa protein, either directly or indirectly, is the product of processing by poliovirus 3CD/3C proteinase. Only the modified 38-kDa protein, not its precursor protein, has a high affinity for binding to the 3' end of viral negative-strand RNA. This modification depends on proteolytically active proteinase, and a direct correlation between the levels of 3CD proteinase and the 38-kDa protein was demonstrated in infected tissue culture cells. The nucleotide (nt) 5-10 region (positive-strand numbers) of poliovirus negative-strand RNA is important for binding of the 38-kDa protein. Deletion of the nt 5-10 region in full-length, positive-strand RNA renders the RNA noninfectious in transfection experiments. These results suggest that poliovirus 3CD/3C proteinase processes a cellular protein which then plays an essential role during the viral life cycle.

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Selected References

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  1. Andino R., Rieckhof G. E., Achacoso P. L., Baltimore D. Poliovirus RNA synthesis utilizes an RNP complex formed around the 5'-end of viral RNA. EMBO J. 1993 Sep;12(9):3587–3598. doi: 10.1002/j.1460-2075.1993.tb06032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andino R., Rieckhof G. E., Baltimore D. A functional ribonucleoprotein complex forms around the 5' end of poliovirus RNA. Cell. 1990 Oct 19;63(2):369–380. doi: 10.1016/0092-8674(90)90170-j. [DOI] [PubMed] [Google Scholar]
  3. Bernstein H. D., Sonenberg N., Baltimore D. Poliovirus mutant that does not selectively inhibit host cell protein synthesis. Mol Cell Biol. 1985 Nov;5(11):2913–2923. doi: 10.1128/mcb.5.11.2913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blair W. S., Li X., Semler B. L. A cellular cofactor facilitates efficient 3CD cleavage of the poliovirus P1 precursor. J Virol. 1993 Apr;67(4):2336–2343. doi: 10.1128/jvi.67.4.2336-2343.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blair W. S., Nguyen J. H., Parsley T. B., Semler B. L. Mutations in the poliovirus 3CD proteinase S1-specificity pocket affect substrate recognition and RNA binding. Virology. 1996 Apr 1;218(1):1–13. doi: 10.1006/viro.1996.0160. [DOI] [PubMed] [Google Scholar]
  6. Blyn L. B., Swiderek K. M., Richards O., Stahl D. C., Semler B. L., Ehrenfeld E. Poly(rC) binding protein 2 binds to stem-loop IV of the poliovirus RNA 5' noncoding region: identification by automated liquid chromatography-tandem mass spectrometry. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):11115–11120. doi: 10.1073/pnas.93.20.11115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Buckley B., Ehrenfeld E. The cap-binding protein complex in uninfected and poliovirus-infected HeLa cells. J Biol Chem. 1987 Oct 5;262(28):13599–13606. [PubMed] [Google Scholar]
  8. Charini W. A., Burns C. C., Ehrenfeld E., Semler B. L. trans rescue of a mutant poliovirus RNA polymerase function. J Virol. 1991 May;65(5):2655–2665. doi: 10.1128/jvi.65.5.2655-2665.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clark M. E., Hämmerle T., Wimmer E., Dasgupta A. Poliovirus proteinase 3C converts an active form of transcription factor IIIC to an inactive form: a mechanism for inhibition of host cell polymerase III transcription by poliovirus. EMBO J. 1991 Oct;10(10):2941–2947. doi: 10.1002/j.1460-2075.1991.tb07844.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clark M. E., Lieberman P. M., Berk A. J., Dasgupta A. Direct cleavage of human TATA-binding protein by poliovirus protease 3C in vivo and in vitro. Mol Cell Biol. 1993 Feb;13(2):1232–1237. doi: 10.1128/mcb.13.2.1232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dewalt P. G., Semler B. L. Site-directed mutagenesis of proteinase 3C results in a poliovirus deficient in synthesis of viral RNA polymerase. J Virol. 1987 Jul;61(7):2162–2170. doi: 10.1128/jvi.61.7.2162-2170.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Etchison D., Milburn S. C., Edery I., Sonenberg N., Hershey J. W. Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220,000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. J Biol Chem. 1982 Dec 25;257(24):14806–14810. [PubMed] [Google Scholar]
  13. Falk M. M., Grigera P. R., Bergmann I. E., Zibert A., Multhaup G., Beck E. Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3. J Virol. 1990 Feb;64(2):748–756. doi: 10.1128/jvi.64.2.748-756.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  15. Furuya T., Lai M. M. Three different cellular proteins bind to complementary sites on the 5'-end-positive and 3'-end-negative strands of mouse hepatitis virus RNA. J Virol. 1993 Dec;67(12):7215–7222. doi: 10.1128/jvi.67.12.7215-7222.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Haller A. A., Nguyen J. H., Semler B. L. Minimum internal ribosome entry site required for poliovirus infectivity. J Virol. 1993 Dec;67(12):7461–7471. doi: 10.1128/jvi.67.12.7461-7471.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Haller A. A., Semler B. L. Linker scanning mutagenesis of the internal ribosome entry site of poliovirus RNA. J Virol. 1992 Aug;66(8):5075–5086. doi: 10.1128/jvi.66.8.5075-5086.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Harris K. S., Xiang W., Alexander L., Lane W. S., Paul A. V., Wimmer E. Interaction of poliovirus polypeptide 3CDpro with the 5' and 3' termini of the poliovirus genome. Identification of viral and cellular cofactors needed for efficient binding. J Biol Chem. 1994 Oct 28;269(43):27004–27014. [PubMed] [Google Scholar]
  19. Joachims M., Harris K. S., Etchison D. Poliovirus protease 3C mediates cleavage of microtubule-associated protein 4. Virology. 1995 Aug 20;211(2):451–461. doi: 10.1006/viro.1995.1427. [DOI] [PubMed] [Google Scholar]
  20. Kräusslich H. G., Nicklin M. J., 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. doi: 10.1128/jvi.61.9.2711-2718.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Käriäinen L., Ranki M. Inhibition of cell functions by RNA-virus infections. Annu Rev Microbiol. 1984;38:91–109. doi: 10.1146/annurev.mi.38.100184.000515. [DOI] [PubMed] [Google Scholar]
  22. La Monica N., Meriam C., Racaniello V. R. Mapping of sequences required for mouse neurovirulence of poliovirus type 2 Lansing. J Virol. 1986 Feb;57(2):515–525. doi: 10.1128/jvi.57.2.515-525.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Leffers H., Dejgaard K., Celis J. E. Characterisation of two major cellular poly(rC)-binding human proteins, each containing three K-homologous (KH) domains. Eur J Biochem. 1995 Jun 1;230(2):447–453. [PubMed] [Google Scholar]
  24. Liebig H. D., Ziegler E., Yan R., Hartmuth K., Klump H., Kowalski H., Blaas D., Sommergruber W., Frasel L., Lamphear B. Purification of two picornaviral 2A proteinases: interaction with eIF-4 gamma and influence on in vitro translation. Biochemistry. 1993 Jul 27;32(29):7581–7588. doi: 10.1021/bi00080a033. [DOI] [PubMed] [Google Scholar]
  25. Lloyd R. E., Grubman M. J., Ehrenfeld E. Relationship of p220 cleavage during picornavirus infection to 2A proteinase sequencing. J Virol. 1988 Nov;62(11):4216–4223. doi: 10.1128/jvi.62.11.4216-4223.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pelletier J., Kaplan G., Racaniello V. R., Sonenberg N. Cap-independent translation of poliovirus mRNA is conferred by sequence elements within the 5' noncoding region. Mol Cell Biol. 1988 Mar;8(3):1103–1112. doi: 10.1128/mcb.8.3.1103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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. doi: 10.1038/334320a0. [DOI] [PubMed] [Google Scholar]
  28. Racaniello V. R., Meriam C. Poliovirus temperature-sensitive mutant containing a single nucleotide deletion in the 5'-noncoding region of the viral RNA. Virology. 1986 Dec;155(2):498–507. doi: 10.1016/0042-6822(86)90211-4. [DOI] [PubMed] [Google Scholar]
  29. Rivera V. M., Welsh J. D., Maizel J. V., Jr Comparative sequence analysis of the 5' noncoding region of the enteroviruses and rhinoviruses. Virology. 1988 Jul;165(1):42–50. doi: 10.1016/0042-6822(88)90656-3. [DOI] [PubMed] [Google Scholar]
  30. Roehl H. H., Anderson M. M., Mehigh C. S., Conrad S. E. Regulation of the cellular thymidine kinase gene promoter in simian virus 40-infected cells. J Virol. 1993 Aug;67(8):4964–4971. doi: 10.1128/jvi.67.8.4964-4971.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Roehl H. H., Semler B. L. Poliovirus infection enhances the formation of two ribonucleoprotein complexes at the 3' end of viral negative-strand RNA. J Virol. 1995 May;69(5):2954–2961. doi: 10.1128/jvi.69.5.2954-2961.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rubinstein S. J., Dasgupta A. Inhibition of rRNA synthesis by poliovirus: specific inactivation of transcription factors. J Virol. 1989 Nov;63(11):4689–4696. doi: 10.1128/jvi.63.11.4689-4696.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sarnow P. Translation of glucose-regulated protein 78/immunoglobulin heavy-chain binding protein mRNA is increased in poliovirus-infected cells at a time when cap-dependent translation of cellular mRNAs is inhibited. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5795–5799. doi: 10.1073/pnas.86.15.5795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Semler B. L., Johnson V. H., Dewalt P. G., Ypma-Wong M. F. Site-specific mutagenesis of cDNA clones expressing a poliovirus proteinase. J Cell Biochem. 1987 Jan;33(1):39–51. doi: 10.1002/jcb.240330105. [DOI] [PubMed] [Google Scholar]
  35. Simoes E. A., 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. doi: 10.1128/jvi.65.2.913-921.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Skinner M. A., Racaniello V. R., Dunn G., Cooper J., Minor P. D., Almond J. W. 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. doi: 10.1016/0022-2836(89)90261-1. [DOI] [PubMed] [Google Scholar]
  37. Tesar M., Marquardt O. Foot-and-mouth disease virus protease 3C inhibits cellular transcription and mediates cleavage of histone H3. Virology. 1990 Feb;174(2):364–374. doi: 10.1016/0042-6822(90)90090-e. [DOI] [PubMed] [Google Scholar]
  38. 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. doi: 10.1128/jvi.62.7.2291-2299.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Walker C., Selby M., Erickson A., Cataldo D., Valensi J. P., Van Nest G. V. Cationic lipids direct a viral glycoprotein into the class I major histocompatibility complex antigen-presentation pathway. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7915–7918. doi: 10.1073/pnas.89.17.7915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Zuker M. On finding all suboptimal foldings of an RNA molecule. Science. 1989 Apr 7;244(4900):48–52. doi: 10.1126/science.2468181. [DOI] [PubMed] [Google Scholar]

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