Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1993 Jun;67(6):3326–3331. doi: 10.1128/jvi.67.6.3326-3331.1993

Identification of the cleavage site and determinants required for poliovirus 3CPro-catalyzed cleavage of human TATA-binding transcription factor TBP.

S Das 1, A Dasgupta 1
PMCID: PMC237675  PMID: 8388502

Abstract

Host cell RNA polymerase II-mediated transcription is inhibited by poliovirus infection. We have shown previously that the human TATA-binding protein (TBP), a general transcription factor required for transcription of all RNA polymerase II genes, is directly cleaved both in vitro and in vivo by the virus-coded protease 3CPro. 3CPro specifically cleaves glutamine-glycine bonds in the viral polyprotein. Cellular transcription factor TBP contains three glutamine-glycine sites, at amino acids 12, 18, and 108. By using site-directed mutagenesis, we determined that the glutamine-glycine bond at amino acid 18, but not that at amino acid 12 or 108, is cleaved by the viral protease. Both the glutamine and the glycine appear to be important for the cleavage. Further mutations around the glutamine-glycine site at position 18 suggest that determinants other than the glutamine-glycine bond in TBP are also required for 3CPro-induced cleavage. An alanine at position P4 and a proline at position P2, proximal to the scissile glutamine-glycine pair, appear to be important for 3CPro-mediated cleavage of TBP. Our results suggest that the cleavage specificity of 3CPro for a cellular transcription factor is very similar to its mode of cleavage of viral polyproteins.

Full text

PDF
3326

Images in this article

3328Image
on p.3328
3328Image
on p.3328
3329Image
on p.3329

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnold E., Luo M., Vriend G., Rossmann M. G., Palmenberg A. C., Parks G. D., Nicklin M. J., Wimmer E. Implications of the picornavirus capsid structure for polyprotein processing. Proc Natl Acad Sci U S A. 1987 Jan;84(1):21–25. doi: 10.1073/pnas.84.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BALTIMORE D., FRANKLIN R. M. The effect of Mengovirus infection on the activity of the DNA-dependent RNA polymerase of L-cells. Proc Natl Acad Sci U S A. 1962 Aug;48:1383–1390. doi: 10.1073/pnas.48.8.1383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blair W. S., Semler B. L. Role for the P4 amino acid residue in substrate utilization by the poliovirus 3CD proteinase. J Virol. 1991 Nov;65(11):6111–6123. doi: 10.1128/jvi.65.11.6111-6123.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buratowski S., Hahn S., Guarente L., Sharp P. A. Five intermediate complexes in transcription initiation by RNA polymerase II. Cell. 1989 Feb 24;56(4):549–561. doi: 10.1016/0092-8674(89)90578-3. [DOI] [PubMed] [Google Scholar]
  5. Clark M. E., Dasgupta A. A transcriptionally active form of TFIIIC is modified in poliovirus-infected HeLa cells. Mol Cell Biol. 1990 Oct;10(10):5106–5113. doi: 10.1128/mcb.10.10.5106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Comai L., Tanese N., Tjian R. The TATA-binding protein and associated factors are integral components of the RNA polymerase I transcription factor, SL1. Cell. 1992 Mar 6;68(5):965–976. doi: 10.1016/0092-8674(92)90039-f. [DOI] [PubMed] [Google Scholar]
  9. Crawford N., Fire A., Samuels M., Sharp P. A., Baltimore D. Inhibition of transcription factor activity by poliovirus. Cell. 1981 Dec;27(3 Pt 2):555–561. doi: 10.1016/0092-8674(81)90397-4. [DOI] [PubMed] [Google Scholar]
  10. Dynlacht B. D., Hoey T., Tjian R. Isolation of coactivators associated with the TATA-binding protein that mediate transcriptional activation. Cell. 1991 Aug 9;66(3):563–576. doi: 10.1016/0092-8674(81)90019-2. [DOI] [PubMed] [Google Scholar]
  11. Hanecak R., Semler B. L., Anderson C. W., Wimmer E. Proteolytic processing of poliovirus polypeptides: antibodies to polypeptide P3-7c inhibit cleavage at glutamine-glycine pairs. Proc Natl Acad Sci U S A. 1982 Jul;79(13):3973–3977. doi: 10.1073/pnas.79.13.3973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harris K. S., Reddigari S. R., Nicklin M. J., Hämmerle T., Wimmer E. Purification and characterization of poliovirus polypeptide 3CD, a proteinase and a precursor for RNA polymerase. J Virol. 1992 Dec;66(12):7481–7489. doi: 10.1128/jvi.66.12.7481-7489.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hellen C. U., Kräusslich H. G., Wimmer E. Proteolytic processing of polyproteins in the replication of RNA viruses. Biochemistry. 1989 Dec 26;28(26):9881–9890. doi: 10.1021/bi00452a001. [DOI] [PubMed] [Google Scholar]
  14. Hoffman A., Sinn E., Yamamoto T., Wang J., Roy A., Horikoshi M., Roeder R. G. Highly conserved core domain and unique N terminus with presumptive regulatory motifs in a human TATA factor (TFIID). Nature. 1990 Jul 26;346(6282):387–390. doi: 10.1038/346387a0. [DOI] [PubMed] [Google Scholar]
  15. Hsiao W. L., Lopez C. A., Wu T., Weinstein I. B. A factor present in fetal calf serum enhances oncogene-induced transformation of rodent fibroblasts. Mol Cell Biol. 1987 Oct;7(10):3380–3385. doi: 10.1128/mcb.7.10.3380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hämmerle T., Hellen C. U., Wimmer E. Site-directed mutagenesis of the putative catalytic triad of poliovirus 3C proteinase. J Biol Chem. 1991 Mar 25;266(9):5412–5416. [PubMed] [Google Scholar]
  17. Kao C. C., Lieberman P. M., Schmidt M. C., Zhou Q., Pei R., Berk A. J. Cloning of a transcriptionally active human TATA binding factor. Science. 1990 Jun 29;248(4963):1646–1650. doi: 10.1126/science.2194289. [DOI] [PubMed] [Google Scholar]
  18. Kliewer S., Dasgupta A. An RNA polymerase II transcription factor inactivated in poliovirus-infected cells copurifies with transcription factor TFIID. Mol Cell Biol. 1988 Aug;8(8):3175–3182. doi: 10.1128/mcb.8.8.3175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kliewer S., Muchardt C., Gaynor R., Dasgupta A. Loss of a phosphorylated form of transcription factor CREB/ATF in poliovirus-infected cells. J Virol. 1990 Sep;64(9):4507–4515. doi: 10.1128/jvi.64.9.4507-4515.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Lawson M. A., Semler B. L. Picornavirus protein processing--enzymes, substrates, and genetic regulation. Curr Top Microbiol Immunol. 1990;161:49–87. [PubMed] [Google Scholar]
  22. Margottin F., Dujardin G., Gérard M., Egly J. M., Huet J., Sentenac A. Participation of the TATA factor in transcription of the yeast U6 gene by RNA polymerase C. Science. 1991 Jan 25;251(4992):424–426. doi: 10.1126/science.1989075. [DOI] [PubMed] [Google Scholar]
  23. Nicklin M. J., Harris K. S., Pallai P. V., Wimmer E. Poliovirus proteinase 3C: large-scale expression, purification, and specific cleavage activity on natural and synthetic substrates in vitro. J Virol. 1988 Dec;62(12):4586–4593. doi: 10.1128/jvi.62.12.4586-4593.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Palmenberg A. C., Kirby E. M., Janda M. R., Drake N. L., Duke G. M., Potratz K. F., Collett M. S. The nucleotide and deduced amino acid sequences of the encephalomyocarditis viral polyprotein coding region. Nucleic Acids Res. 1984 Mar 26;12(6):2969–2985. doi: 10.1093/nar/12.6.2969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Palmenberg A. C. Proteolytic processing of picornaviral polyprotein. Annu Rev Microbiol. 1990;44:603–623. doi: 10.1146/annurev.mi.44.100190.003131. [DOI] [PubMed] [Google Scholar]
  26. Parks G. D., Baker J. C., Palmenberg A. C. Proteolytic cleavage of encephalomyocarditis virus capsid region substrates by precursors to the 3C enzyme. J Virol. 1989 Mar;63(3):1054–1058. doi: 10.1128/jvi.63.3.1054-1058.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Peterson M. G., Tanese N., Pugh B. F., Tjian R. Functional domains and upstream activation properties of cloned human TATA binding protein. Science. 1990 Jun 29;248(4963):1625–1630. doi: 10.1126/science.2363050. [DOI] [PubMed] [Google Scholar]
  28. Pugh B. F., Tjian R. Diverse transcriptional functions of the multisubunit eukaryotic TFIID complex. J Biol Chem. 1992 Jan 15;267(2):679–682. [PubMed] [Google Scholar]
  29. Pugh B. F., Tjian R. Transcription from a TATA-less promoter requires a multisubunit TFIID complex. Genes Dev. 1991 Nov;5(11):1935–1945. doi: 10.1101/gad.5.11.1935. [DOI] [PubMed] [Google Scholar]
  30. Roeder R. G. The complexities of eukaryotic transcription initiation: regulation of preinitiation complex assembly. Trends Biochem Sci. 1991 Nov;16(11):402–408. doi: 10.1016/0968-0004(91)90164-q. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Rubinstein S. J., Hammerle T., Wimmer E., Dasgupta A. Infection of HeLa cells with poliovirus results in modification of a complex that binds to the rRNA promoter. J Virol. 1992 May;66(5):3062–3068. doi: 10.1128/jvi.66.5.3062-3068.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sharp P. A. TATA-binding protein is a classless factor. Cell. 1992 Mar 6;68(5):819–821. doi: 10.1016/0092-8674(92)90023-6. [DOI] [PubMed] [Google Scholar]
  35. Van Dyke M. W., Roeder R. G., Sawadogo M. Physical analysis of transcription preinitiation complex assembly on a class II gene promoter. Science. 1988 Sep 9;241(4871):1335–1338. doi: 10.1126/science.3413495. [DOI] [PubMed] [Google Scholar]
  36. Ypma-Wong M. F., Dewalt P. G., Johnson V. H., Lamb J. G., Semler B. L. Protein 3CD is the major poliovirus proteinase responsible for cleavage of the P1 capsid precursor. Virology. 1988 Sep;166(1):265–270. doi: 10.1016/0042-6822(88)90172-9. [DOI] [PubMed] [Google Scholar]
  37. Ypma-Wong M. F., Filman D. J., Hogle J. M., Semler B. L. Structural domains of the poliovirus polyprotein are major determinants for proteolytic cleavage at Gln-Gly pairs. J Biol Chem. 1988 Nov 25;263(33):17846–17856. [PubMed] [Google Scholar]
  38. Zawel L., Reinberg D. Advances in RNA polymerase II transcription. Curr Opin Cell Biol. 1992 Jun;4(3):488–495. doi: 10.1016/0955-0674(92)90016-6. [DOI] [PubMed] [Google Scholar]
  39. Zhou Q., Lieberman P. M., Boyer T. G., Berk A. J. Holo-TFIID supports transcriptional stimulation by diverse activators and from a TATA-less promoter. Genes Dev. 1992 Oct;6(10):1964–1974. doi: 10.1101/gad.6.10.1964. [DOI] [PubMed] [Google Scholar]

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

RESOURCES