Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1996 Aug;16(8):4163–4171. doi: 10.1128/mcb.16.8.4163

DNA binding domain and subunit interactions of transcription factor IIIC revealed by dissection with poliovirus 3C protease.

Y Shen 1, M Igo 1, P Yalamanchili 1, A J Berk 1, A Dasgupta 1
PMCID: PMC231413  PMID: 8754815

Abstract

Transcription factor IIIC (TFIIIC) is a general RNA polymerase III transcription factor that binds the B-box internal promotor element of tRNA genes and the complex of TFIIIA with a 5S rRNA gene. TFIIIC then directs the binding of TFIIIB to DNA upstream of the transcription start site. TFIIIB in turn directs RNA polymerase III binding and initiation. Human TFIIIC contains five different subunits. The 243-kDa alpha subunit can be specifically cross-linked to B-box DNA, but its sequence does not reveal a known DNA binding domain. During poliovirus infection, TFIIIC is cleaved and inactivated by the poliovirus-encoded 3C protease (3Cpro). Here we analyzed the cleavage of TFIIIC subunits by 3Cpro in vitro and during poliovirus infection of HeLa cells. Analyses of the DNA binding activities of the resulting subcomplexes indicated that an N-terminal 83-kDa domain of the alpha subunit associates with the beta subunit to generate the TFIIIC DNA binding domain. Cleavage with 3Cpro also generated an approximately 125-kDa C-terminal fragment of the alpha subunit which remained associated with the gamma and epsilon subunits.

Full Text

The Full Text of this article is available as a PDF (814.1 KB).

Selected References

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

  1. Balanian R. Structural and functional alterations in cultured cells infected with cytocidal viruses. Prog Med Virol. 1975;19:40–83. [PubMed] [Google Scholar]
  2. Bartholomew B., Braun B. R., Kassavetis G. A., Geiduschek E. P. Probing close DNA contacts of RNA polymerase III transcription complexes with the photoactive nucleoside 4-thiodeoxythymidine. J Biol Chem. 1994 Jul 8;269(27):18090–18095. [PubMed] [Google Scholar]
  3. Bartholomew B., Kassavetis G. A., Geiduschek E. P. Two components of Saccharomyces cerevisiae transcription factor IIIB (TFIIIB) are stereospecifically located upstream of a tRNA gene and interact with the second-largest subunit of TFIIIC. Mol Cell Biol. 1991 Oct;11(10):5181–5189. doi: 10.1128/mcb.11.10.5181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boulanger P. A., L'Etoile N. D., Berk A. J. A DNA-binding domain of human transcription factor IIIC2. Nucleic Acids Res. 1989 Oct 11;17(19):7761–7770. doi: 10.1093/nar/17.19.7761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chaussivert N., Conesa C., Shaaban S., Sentenac A. Complex interactions between yeast TFIIIB and TFIIIC. J Biol Chem. 1995 Jun 23;270(25):15353–15358. doi: 10.1074/jbc.270.25.15353. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. 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]
  9. Conesa C., Swanson R. N., Schultz P., Oudet P., Sentenac A. On the subunit composition, stoichiometry, and phosphorylation of the yeast transcription factor TFIIIC/tau. J Biol Chem. 1993 Aug 25;268(24):18047–18052. [PubMed] [Google Scholar]
  10. 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]
  11. 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. doi: 10.1128/jvi.67.6.3326-3331.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dasgupta A. Purification of host factor required for in vitro transcription of poliovirus RNA. Virology. 1983 Jul 15;128(1):245–251. doi: 10.1016/0042-6822(83)90335-5. [DOI] [PubMed] [Google Scholar]
  13. Dignam J. D., Lebovitz R. M., Roeder R. G. 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. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Elroy-Stein O., Fuerst T. R., Moss B. Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6126–6130. doi: 10.1073/pnas.86.16.6126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fradkin L. G., Yoshinaga S. K., Berk A. J., Dasgupta A. Inhibition of host cell RNA polymerase III-mediated transcription by poliovirus: inactivation of specific transcription factors. Mol Cell Biol. 1987 Nov;7(11):3880–3887. doi: 10.1128/mcb.7.11.3880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gabrielsen O. S., Marzouki N., Ruet A., Sentenac A., Fromageot P. Two polypeptide chains in yeast transcription factor tau interact with DNA. J Biol Chem. 1989 May 5;264(13):7505–7511. [PubMed] [Google Scholar]
  17. Geiduschek E. P., Kassavetis G. A. Comparing transcriptional initiation by RNA polymerases I and III. Curr Opin Cell Biol. 1995 Jun;7(3):344–351. doi: 10.1016/0955-0674(95)80089-1. [DOI] [PubMed] [Google Scholar]
  18. Kassavetis G. A., Braun B. R., Nguyen L. H., Geiduschek E. P. S. cerevisiae TFIIIB is the transcription initiation factor proper of RNA polymerase III, while TFIIIA and TFIIIC are assembly factors. Cell. 1990 Jan 26;60(2):235–245. doi: 10.1016/0092-8674(90)90739-2. [DOI] [PubMed] [Google Scholar]
  19. Khoo B., Brophy B., Jackson S. P. Conserved functional domains of the RNA polymerase III general transcription factor BRF. Genes Dev. 1994 Dec 1;8(23):2879–2890. doi: 10.1101/gad.8.23.2879. [DOI] [PubMed] [Google Scholar]
  20. Kovelman R., Roeder R. G. Purification and characterization of two forms of human transcription factor IIIC. J Biol Chem. 1992 Dec 5;267(34):24446–24456. [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. L'Etoile N. D., Fahnestock M. L., Shen Y., Aebersold R., Berk A. J. Human transcription factor IIIC box B binding subunit. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1652–1656. doi: 10.1073/pnas.91.5.1652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lagna G., Kovelman R., Sukegawa J., Roeder R. G. Cloning and characterization of an evolutionarily divergent DNA-binding subunit of mammalian TFIIIC. Mol Cell Biol. 1994 May;14(5):3053–3064. doi: 10.1128/mcb.14.5.3053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lee K. A., Green M. R. Small-scale preparation of extracts from radiolabeled cells efficient in pre-mRNA splicing. Methods Enzymol. 1990;181:20–30. doi: 10.1016/0076-6879(90)81108-7. [DOI] [PubMed] [Google Scholar]
  25. Lefebvre O., Carles C., Conesa C., Swanson R. N., Bouet F., Riva M., Sentenac A. TFC3: gene encoding the B-block binding subunit of the yeast transcription factor IIIC. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10512–10516. doi: 10.1073/pnas.89.21.10512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marck C., Lefebvre O., Carles C., Riva M., Chaussivert N., Ruet A., Sentenac A. The TFIIIB-assembling subunit of yeast transcription factor TFIIIC has both tetratricopeptide repeats and basic helix-loop-helix motifs. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4027–4031. doi: 10.1073/pnas.90.9.4027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Schwartz L. B., Lawrence C., Thach R. E., Roeder R. G. Encephalomyocarditis virus infection of mouse plasmacytoma cells. II. Effect on host RNA synthesis and RNA polymerases. J Virol. 1974 Sep;14(3):611–619. doi: 10.1128/jvi.14.3.611-619.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sinn E., Wang Z., Kovelman R., Roeder R. G. Cloning and characterization of a TFIIIC2 subunit (TFIIIC beta) whose presence correlates with activation of RNA polymerase III-mediated transcription by adenovirus E1A expression and serum factors. Genes Dev. 1995 Mar 15;9(6):675–685. doi: 10.1101/gad.9.6.675. [DOI] [PubMed] [Google Scholar]
  31. Werner M., Chaussivert N., Willis I. M., Sentenac A. Interaction between a complex of RNA polymerase III subunits and the 70-kDa component of transcription factor IIIB. J Biol Chem. 1993 Oct 5;268(28):20721–20724. [PubMed] [Google Scholar]
  32. Willis I. M. RNA polymerase III. Genes, factors and transcriptional specificity. Eur J Biochem. 1993 Feb 15;212(1):1–11. doi: 10.1111/j.1432-1033.1993.tb17626.x. [DOI] [PubMed] [Google Scholar]
  33. Yoshinaga S. K., Boulanger P. A., Berk A. J. Resolution of human transcription factor TFIIIC into two functional components. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3585–3589. doi: 10.1073/pnas.84.11.3585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yoshinaga S. K., L'Etoile N. D., Berk A. J. Purification and characterization of transcription factor IIIC2. J Biol Chem. 1989 Jun 25;264(18):10726–10731. [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES