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. 1997 Sep;17(9):5299–5306. doi: 10.1128/mcb.17.9.5299

Domains of the Brf component of RNA polymerase III transcription factor IIIB (TFIIIB): functions in assembly of TFIIIB-DNA complexes and recruitment of RNA polymerase to the promoter.

G A Kassavetis 1, C Bardeleben 1, A Kumar 1, E Ramirez 1, E P Geiduschek 1
PMCID: PMC232380  PMID: 9271407

Abstract

Saccharomyces cerevisiae transcription factor IIIB (TFIIIB) is composed of three subunits: the TATA-binding protein, the TFIIB-related protein Brf, and B". TFIIIB, which is brought to RNA polymerase III-transcribed genes indirectly through interaction with DNA-bound TFIIIC or directly through DNA recognition by the TATA-binding protein, in turn recruits RNA polymerase III to the promoter. N-terminally deleted derivatives of Brf have been examined for their ability to interact with DNA-bound TFIIIC and with the other components of TFIIIB and for participation in transcription. Brf(165-596), lacking 164 N-proximal TFIIB-homologous amino acids, is competent to participate in the assembly of TFIIIB-DNA complexes and in TFIIIC-independent transcription. Even deletion of the entire TFIIB-homologous half of the protein, as in Brf(317-596) and Brf(352-596), allows some interaction with DNA-bound TBP and with the B" component of TFIIIB to be retained. The function of Brf(165-596) in transcription has also been examined in the context of B" with small internal deletions. The ability of Brf with this sizable N-terminal segment deleted to function in TFIIIC-independent transcription requires segments of B" that are individually indispensable although required on an either/or basis, in the context of complete Brf. These findings suggest a functional complementarity and reciprocity between the Brf and B" components of TFIIIB.

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Selected References

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  1. Baker R. E., Camier S., Sentenac A., Hall B. D. Gene size differentially affects the binding of yeast transcription factor tau to two intragenic regions. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8768–8772. doi: 10.1073/pnas.84.24.8768. [DOI] [PMC free article] [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., Durkovich D., Kassavetis G. A., Geiduschek E. P. Orientation and topography of RNA polymerase III in transcription complexes. Mol Cell Biol. 1993 Feb;13(2):942–952. doi: 10.1128/mcb.13.2.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bartholomew B., Kassavetis G. A., Braun B. R., Geiduschek E. P. The subunit structure of Saccharomyces cerevisiae transcription factor IIIC probed with a novel photocrosslinking reagent. EMBO J. 1990 Jul;9(7):2197–2205. doi: 10.1002/j.1460-2075.1990.tb07389.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Bartholomew B., Tinker R. L., Kassavetis G. A., Geiduschek E. P. Photochemical cross-linking assay for DNA tracking by replication proteins. Methods Enzymol. 1995;262:476–494. doi: 10.1016/0076-6879(95)62039-7. [DOI] [PubMed] [Google Scholar]
  7. Braun B. R., Riggs D. L., Kassavetis G. A., Geiduschek E. P. Multiple states of protein-DNA interaction in the assembly of transcription complexes on Saccharomyces cerevisiae 5S ribosomal RNA genes. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2530–2534. doi: 10.1073/pnas.86.8.2530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buratowski S., Zhou H. A suppressor of TBP mutations encodes an RNA polymerase III transcription factor with homology to TFIIB. Cell. 1992 Oct 16;71(2):221–230. doi: 10.1016/0092-8674(92)90351-c. [DOI] [PubMed] [Google Scholar]
  9. Burley S. K., Roeder R. G. Biochemistry and structural biology of transcription factor IID (TFIID). Annu Rev Biochem. 1996;65:769–799. doi: 10.1146/annurev.bi.65.070196.004005. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Chiang C. M., Ge H., Wang Z., Hoffmann A., Roeder R. G. Unique TATA-binding protein-containing complexes and cofactors involved in transcription by RNA polymerases II and III. EMBO J. 1993 Jul;12(7):2749–2762. doi: 10.1002/j.1460-2075.1993.tb05936.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Colbert T., Hahn S. A yeast TFIIB-related factor involved in RNA polymerase III transcription. Genes Dev. 1992 Oct;6(10):1940–1949. doi: 10.1101/gad.6.10.1940. [DOI] [PubMed] [Google Scholar]
  13. Dieci G., Duimio L., Coda-Zabetta F., Sprague K. U., Ottonello S. A novel RNA polymerase III transcription factor fraction that is not required for template commitment. J Biol Chem. 1993 May 25;268(15):11199–11207. [PubMed] [Google Scholar]
  14. Eschenlauer J. B., Kaiser M. W., Gerlach V. L., Brow D. A. Architecture of a yeast U6 RNA gene promoter. Mol Cell Biol. 1993 May;13(5):3015–3026. doi: 10.1128/mcb.13.5.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Geiduschek E. P., Tocchini-Valentini G. P. Transcription by RNA polymerase III. Annu Rev Biochem. 1988;57:873–914. doi: 10.1146/annurev.bi.57.070188.004301. [DOI] [PubMed] [Google Scholar]
  17. Gerlach V. L., Whitehall S. K., Geiduschek E. P., Brow D. A. TFIIIB placement on a yeast U6 RNA gene in vivo is directed primarily by TFIIIC rather than by sequence-specific DNA contacts. Mol Cell Biol. 1995 Mar;15(3):1455–1466. doi: 10.1128/mcb.15.3.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ha I., Roberts S., Maldonado E., Sun X., Kim L. U., Green M., Reinberg D. Multiple functional domains of human transcription factor IIB: distinct interactions with two general transcription factors and RNA polymerase II. Genes Dev. 1993 Jun;7(6):1021–1032. doi: 10.1101/gad.7.6.1021. [DOI] [PubMed] [Google Scholar]
  19. Hager D. A., Burgess R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 1980 Nov 15;109(1):76–86. doi: 10.1016/0003-2697(80)90013-5. [DOI] [PubMed] [Google Scholar]
  20. Hisatake K., Roeder R. G., Horikoshi M. Functional dissection of TFIIB domains required for TFIIB-TFIID-promoter complex formation and basal transcription activity. Nature. 1993 Jun 24;363(6431):744–747. doi: 10.1038/363744a0. [DOI] [PubMed] [Google Scholar]
  21. Huet J., Conesa C., Manaud N., Chaussivert N., Sentenac A. Interactions between yeast TFIIIB components. Nucleic Acids Res. 1994 Aug 25;22(16):3433–3439. doi: 10.1093/nar/22.16.3433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kassavetis G. A., Bartholomew B., Blanco J. A., Johnson T. E., Geiduschek E. P. Two essential components of the Saccharomyces cerevisiae transcription factor TFIIIB: transcription and DNA-binding properties. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7308–7312. doi: 10.1073/pnas.88.16.7308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Kassavetis G. A., Joazeiro C. A., Pisano M., Geiduschek E. P., Colbert T., Hahn S., Blanco J. A. The role of the TATA-binding protein in the assembly and function of the multisubunit yeast RNA polymerase III transcription factor, TFIIIB. Cell. 1992 Dec 11;71(6):1055–1064. doi: 10.1016/0092-8674(92)90399-w. [DOI] [PubMed] [Google Scholar]
  25. Kassavetis G. A., Nguyen S. T., Kobayashi R., Kumar A., Geiduschek E. P., Pisano M. Cloning, expression, and function of TFC5, the gene encoding the B" component of the Saccharomyces cerevisiae RNA polymerase III transcription factor TFIIIB. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9786–9790. doi: 10.1073/pnas.92.21.9786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Kumar A., Kassavetis G. A., Geiduschek E. P., Hambalko M., Brent C. J. Functional dissection of the B" component of RNA polymerase III transcription factor IIIB: a scaffolding protein with multiple roles in assembly and initiation of transcription. Mol Cell Biol. 1997 Apr;17(4):1868–1880. doi: 10.1128/mcb.17.4.1868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  29. Lagrange T., Kim T. K., Orphanides G., Ebright Y. W., Ebright R. H., Reinberg D. High-resolution mapping of nucleoprotein complexes by site-specific protein-DNA photocrosslinking: organization of the human TBP-TFIIA-TFIIB-DNA quaternary complex. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10620–10625. doi: 10.1073/pnas.93.20.10620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Librizzi M. D., Moir R. D., Brenowitz M., Willis I. M. Expression and purification of the RNA polymerase III transcription specificity factor IIIB70 from Saccharomyces cerevisiae and its cooperative binding with TATA-binding protein. J Biol Chem. 1996 Dec 20;271(51):32695–32701. doi: 10.1074/jbc.271.51.32695. [DOI] [PubMed] [Google Scholar]
  31. Lobo S. M., Tanaka M., Sullivan M. L., Hernandez N. A TBP complex essential for transcription from TATA-less but not TATA-containing RNA polymerase III promoters is part of the TFIIIB fraction. Cell. 1992 Dec 11;71(6):1029–1040. doi: 10.1016/0092-8674(92)90397-u. [DOI] [PubMed] [Google Scholar]
  32. Léveillard T., Kassavetis G. A., Geiduschek E. P. Saccharomyces cerevisiae transcription factors IIIB and IIIC bend the DNA of a tRNA(Gln) gene. J Biol Chem. 1991 Mar 15;266(8):5162–5168. [PubMed] [Google Scholar]
  33. López-De-León A., Librizzi M., Puglia K., Willis I. M. PCF4 encodes an RNA polymerase III transcription factor with homology to TFIIB. Cell. 1992 Oct 16;71(2):211–220. doi: 10.1016/0092-8674(92)90350-l. [DOI] [PubMed] [Google Scholar]
  34. Mital R., Kobayashi R., Hernandez N. RNA polymerase III transcription from the human U6 and adenovirus type 2 VAI promoters has different requirements for human BRF, a subunit of human TFIIIB. Mol Cell Biol. 1996 Dec;16(12):7031–7042. doi: 10.1128/mcb.16.12.7031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nikolov D. B., Chen H., Halay E. D., Usheva A. A., Hisatake K., Lee D. K., Roeder R. G., Burley S. K. Crystal structure of a TFIIB-TBP-TATA-element ternary complex. Nature. 1995 Sep 14;377(6545):119–128. doi: 10.1038/377119a0. [DOI] [PubMed] [Google Scholar]
  36. Poon D., Weil P. A. Immunopurification of yeast TATA-binding protein and associated factors. Presence of transcription factor IIIB transcriptional activity. J Biol Chem. 1993 Jul 25;268(21):15325–15328. [PubMed] [Google Scholar]
  37. Roberts S., Miller S. J., Lane W. S., Lee S., Hahn S. Cloning and functional characterization of the gene encoding the TFIIIB90 subunit of RNA polymerase III transcription factor TFIIIB. J Biol Chem. 1996 Jun 21;271(25):14903–14909. doi: 10.1074/jbc.271.25.14903. [DOI] [PubMed] [Google Scholar]
  38. Rüth J., Conesa C., Dieci G., Lefebvre O., Düsterhöft A., Ottonello S., Sentenac A. A suppressor of mutations in the class III transcription system encodes a component of yeast TFIIIB. EMBO J. 1996 Apr 15;15(8):1941–1949. [PMC free article] [PubMed] [Google Scholar]
  39. Taggart A. K., Fisher T. S., Pugh B. F. The TATA-binding protein and associated factors are components of pol III transcription factor TFIIIB. Cell. 1992 Dec 11;71(6):1015–1028. doi: 10.1016/0092-8674(92)90396-t. [DOI] [PubMed] [Google Scholar]
  40. Wang Z., Roeder R. G. Structure and function of a human transcription factor TFIIIB subunit that is evolutionarily conserved and contains both TFIIB- and high-mobility-group protein 2-related domains. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7026–7030. doi: 10.1073/pnas.92.15.7026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. Whitehall S. K., Kassavetis G. A., Geiduschek E. P. The symmetry of the yeast U6 RNA gene's TATA box and the orientation of the TATA-binding protein in yeast TFIIIB. Genes Dev. 1995 Dec 1;9(23):2974–2985. doi: 10.1101/gad.9.23.2974. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Yamashita S., Hisatake K., Kokubo T., Doi K., Roeder R. G., Horikoshi M., Nakatani Y. Transcription factor TFIIB sites important for interaction with promoter-bound TFIID. Science. 1993 Jul 23;261(5120):463–466. doi: 10.1126/science.8332911. [DOI] [PubMed] [Google Scholar]

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