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J Virol. May 1996; 70(5): 2922–2929.
PMCID: PMC190150

Inhibition of basal transcription by poliovirus: a virus- encoded protease (3Cpro) inhibits formation of TBP-TATA box complex in vitro.


Host cell RNA polymerase II (pol II)-mediated transcription is inhibited by poliovirus infection. We demonstrate here that both TATA- and initiator-mediated basal transcription is inhibited in extracts prepared from poliovirus-infected HeLa cells. This inhibition can be reproduced by incubation of uninfected HeLa cell extracts with purified, recombinant poliovirus protease, 3Cpro. Transient-transfection assays demonstrate that 3Cpro, in the absence of other viral proteins, is able to inhibit cellular pol II-mediated transcription in vivo. Three lines of evidence suggest that inactivation of TATA-binding protein (TBP) is the major cause of inhibition of basal transcription by poliovirus. First, RNA pol II transcription in poliovirus-infected cell extract is fully restored by bacterially expressed TBP. Second, addition of purified TBP restores transcription in heat-treated nuclear extracts from mock- and virus-infected cells to identical levels. Finally, using a gel mobility shift assay, we demonstrate that incubation of TBP with the viral protease (3Cpro) inhibits its ability to bind TATA sequence in vitro. These results suggest that inhibition of pol II transcription in mammalian cells infected with poliovirus is, at least in part, due to the inability of modified TBP to bind pol II promoter sequences.

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Selected References

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  • Apriletti JW, Penhoet EE. Cellular RNA synthesis in normal and mengovirus-infected L-929 cells. J Biol Chem. 1978 Jan 25;253(2):603–611. [PubMed]
  • Balanian R. Structural and functional alterations in cultured cells infected with cytocidal viruses. Prog Med Virol. 1975;19:40–83. [PubMed]
  • BALTIMORE D, FRANKLIN RM. 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. [PMC free article] [PubMed]
  • Basu A, Park K, Atchison ML, Carter RS, Avadhani NG. Identification of a transcriptional initiator element in the cytochrome c oxidase subunit Vb promoter which binds to transcription factors NF-E1 (YY-1, delta) and Sp1. J Biol Chem. 1993 Feb 25;268(6):4188–4196. [PubMed]
  • Breathnach R, Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. [PubMed]
  • Buermeyer AB, Thompson NE, Strasheim LA, Burgess RR, Farnham PJ. The HIP1 initiator element plays a role in determining the in vitro requirement of the dihydrofolate reductase gene promoter for the C-terminal domain of RNA polymerase II. Mol Cell Biol. 1992 May;12(5):2250–2259. [PMC free article] [PubMed]
  • Carcamo J, Buckbinder L, Reinberg D. The initiator directs the assembly of a transcription factor IID-dependent transcription complex. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8052–8056. [PMC free article] [PubMed]
  • Clark ME, Dasgupta A. A transcriptionally active form of TFIIIC is modified in poliovirus-infected HeLa cells. Mol Cell Biol. 1990 Oct;10(10):5106–5113. [PMC free article] [PubMed]
  • Clark ME, 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. [PMC free article] [PubMed]
  • Clark ME, Lieberman PM, Berk AJ, 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. [PMC free article] [PubMed]
  • Cortes P, Flores O, Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II: purification and analysis of transcription factor IIA and identification of transcription factor IIJ. Mol Cell Biol. 1992 Jan;12(1):413–421. [PMC free article] [PubMed]
  • Crawford N, Fire A, Samuels M, Sharp PA, Baltimore D. Inhibition of transcription factor activity by poliovirus. Cell. 1981 Dec;27(3 Pt 2):555–561. [PubMed]
  • Das S, Dasgupta A. Identification of the cleavage site and determinants required for poliovirus 3CPro-catalyzed cleavage of human TATA-binding transcription factor TBP. J Virol. 1993 Jun;67(6):3326–3331. [PMC free article] [PubMed]
  • Dewalt PG, Semler BL. 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. [PMC free article] [PubMed]
  • Dignam JD, Lebovitz RM, Roeder RG. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. [PMC free article] [PubMed]
  • Dynan WS, Tjian R. Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. Nature. 316(6031):774–778. [PubMed]
  • Fernández-Tomás C. The presence of viral-induced proteins in nuclei from poliovirus-infected HeLa cells. Virology. 1982 Jan 30;116(2):629–634. [PubMed]
  • Flores O, Lu H, Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II. Identification and characterization of factor IIH. J Biol Chem. 1992 Feb 5;267(4):2786–2793. [PubMed]
  • Hsiao WL, Lopez CA, Wu T, Weinstein IB. A factor present in fetal calf serum enhances oncogene-induced transformation of rodent fibroblasts. Mol Cell Biol. 1987 Oct;7(10):3380–3385. [PMC free article] [PubMed]
  • Hämmerle T, Hellen CU, 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]
  • Javahery R, Khachi A, Lo K, Zenzie-Gregory B, Smale ST. DNA sequence requirements for transcriptional initiator activity in mammalian cells. Mol Cell Biol. 1994 Jan;14(1):116–127. [PMC free article] [PubMed]
  • Käriäinen L, Ranki M. Inhibition of cell functions by RNA-virus infections. Annu Rev Microbiol. 1984;38:91–109. [PubMed]
  • Kao CC, Lieberman PM, Schmidt MC, Zhou Q, Pei R, Berk AJ. Cloning of a transcriptionally active human TATA binding factor. Science. 1990 Jun 29;248(4963):1646–1650. [PubMed]
  • 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. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Kollmar R, Farnham PJ. Site-specific initiation of transcription by RNA polymerase II. Proc Soc Exp Biol Med. 1993 Jun;203(2):127–139. [PubMed]
  • Nicklin MJ, Harris KS, Pallai PV, 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. [PMC free article] [PubMed]
  • Peterson MG, Tanese N, Pugh BF, Tjian R. Functional domains and upstream activation properties of cloned human TATA binding protein. Science. 1990 Jun 29;248(4963):1625–1630. [PubMed]
  • Pugh BF, Tjian R. Transcription from a TATA-less promoter requires a multisubunit TFIID complex. Genes Dev. 1991 Nov;5(11):1935–1945. [PubMed]
  • Reinberg D, Horikoshi M, Roeder RG. Factors involved in specific transcription in mammalian RNA polymerase II. Functional analysis of initiation factors IIA and IID and identification of a new factor operating at sequences downstream of the initiation site. J Biol Chem. 1987 Mar 5;262(7):3322–3330. [PubMed]
  • Reinberg D, Roeder RG. Factors involved in specific transcription by mammalian RNA polymerase II. Purification and functional analysis of initiation factors IIB and IIE. J Biol Chem. 1987 Mar 5;262(7):3310–3321. [PubMed]
  • Roeder RG. The complexities of eukaryotic transcription initiation: regulation of preinitiation complex assembly. Trends Biochem Sci. 1991 Nov;16(11):402–408. [PubMed]
  • Roy AL, Meisterernst M, Pognonec P, Roeder RG. Cooperative interaction of an initiator-binding transcription initiation factor and the helix-loop-helix activator USF. Nature. 1991 Nov 21;354(6350):245–248. [PubMed]
  • Rubinstein SJ, 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. [PMC free article] [PubMed]
  • Sawadogo M, Roeder RG. Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region. Cell. 1985 Nov;43(1):165–175. [PubMed]
  • Sawadogo M, Sentenac A. RNA polymerase B (II) and general transcription factors. Annu Rev Biochem. 1990;59:711–754. [PubMed]
  • Schwartz LB, Lawrence C, Thach RE, Roeder RG. Encephalomyocarditis virus infection of mouse plasmacytoma cells. II. Effect on host RNA synthesis and RNA polymerases. J Virol. 1974 Sep;14(3):611–619. [PMC free article] [PubMed]
  • Seto E, Shi Y, Shenk T. YY1 is an initiator sequence-binding protein that directs and activates transcription in vitro. Nature. 1991 Nov 21;354(6350):241–245. [PubMed]
  • Smale ST, Baltimore D. The "initiator" as a transcription control element. Cell. 1989 Apr 7;57(1):103–113. [PubMed]
  • Smale ST, Schmidt MC, Berk AJ, Baltimore D. Transcriptional activation by Sp1 as directed through TATA or initiator: specific requirement for mammalian transcription factor IID. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4509–4513. [PMC free article] [PubMed]
  • Urzainqui A, Carrasco L. Degradation of cellular proteins during poliovirus infection: studies by two-dimensional gel electrophoresis. J Virol. 1989 Nov;63(11):4729–4735. [PMC free article] [PubMed]
  • Weis L, Reinberg D. Transcription by RNA polymerase II: initiator-directed formation of transcription-competent complexes. FASEB J. 1992 Nov;6(14):3300–3309. [PubMed]
  • Workman JL, Roeder RG. Binding of transcription factor TFIID to the major late promoter during in vitro nucleosome assembly potentiates subsequent initiation by RNA polymerase II. Cell. 1987 Nov 20;51(4):613–622. [PubMed]
  • Zawel L, Reinberg D. Advances in RNA polymerase II transcription. Curr Opin Cell Biol. 1992 Jun;4(3):488–495. [PubMed]
  • Zenzie-Gregory B, Khachi A, Garraway IP, Smale ST. Mechanism of initiator-mediated transcription: evidence for a functional interaction between the TATA-binding protein and DNA in the absence of a specific recognition sequence. Mol Cell Biol. 1993 Jul;13(7):3841–3849. [PMC free article] [PubMed]
  • Zhou Q, Lieberman PM, Boyer TG, Berk AJ. Holo-TFIID supports transcriptional stimulation by diverse activators and from a TATA-less promoter. Genes Dev. 1992 Oct;6(10):1964–1974. [PubMed]

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