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. 1993 Feb 25;21(4):1013–1018. doi: 10.1093/nar/21.4.1013

Transcription factor IIA stimulates the expression of classical polIII-genes.

W Meissner 1, R Holland 1, R Waldschmidt 1, K H Seifart 1
PMCID: PMC309237  PMID: 8451168

Abstract

Protein fractions containing TFIIA, a transcription factor known to be involved in transcription initiation by RNA polymerase II and 5'-regulated polymerase III genes (e.g. U6), were tested for their role in in vitro transcription of classical pol III genes. These fractions were shown to stimulate a basal transcription system, reconstituted from highly purified fractions hTFIIIB and hTFIIIC. We demonstrate that this stimulating activity isolated from HeLa cells coelutes over at least six chromatographic steps with hTFIIA. Moreover the native molecular mass and the stability of this activity against heat treatment are comparable to those of hTFIIA. Finally we show that recombinant TFIIA from Saccharomyces cerevisiae can substitute for the human factor in pol III transcription in vitro which proves that TFIIA is also involved in the efficient expression of classical pol III genes.

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Selected References

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

  1. Bungert J., Waldschmidt R., Kober I., Seifart K. H. Transcription factor IIA is inactivated during terminal differentiation of avian erythroid cells. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11678–11682. doi: 10.1073/pnas.89.24.11678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Cormack B. P., Struhl K. The TATA-binding protein is required for transcription by all three nuclear RNA polymerases in yeast cells. Cell. 1992 May 15;69(4):685–696. doi: 10.1016/0092-8674(92)90232-2. [DOI] [PubMed] [Google Scholar]
  4. Egly J. M., Miyamoto N. G., Moncollin V., Chambon P. Is actin a transcription initiation factor for RNA polymerase B? EMBO J. 1984 Oct;3(10):2363–2371. doi: 10.1002/j.1460-2075.1984.tb02141.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gabrielsen O. S., Sentenac A. RNA polymerase III (C) and its transcription factors. Trends Biochem Sci. 1991 Nov;16(11):412–416. doi: 10.1016/0968-0004(91)90166-s. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Huet J., Sentenac A. The TATA-binding protein participates in TFIIIB assembly on tRNA genes. Nucleic Acids Res. 1992 Dec 25;20(24):6451–6454. doi: 10.1093/nar/20.24.6451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ilag L. L., Jahn D., Eggertsson G., Söll D. The Escherichia coli hemL gene encodes glutamate 1-semialdehyde aminotransferase. J Bacteriol. 1991 Jun;173(11):3408–3413. doi: 10.1128/jb.173.11.3408-3413.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jahn D., Wingender E., Seifart K. H. Transcription complexes for various class III genes differ in parameters of formation and stability towards salt. J Mol Biol. 1987 Jan 20;193(2):303–313. doi: 10.1016/0022-2836(87)90221-x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Lee D. K., DeJong J., Hashimoto S., Horikoshi M., Roeder R. G. TFIIA induces conformational changes in TFIID via interactions with the basic repeat. Mol Cell Biol. 1992 Nov;12(11):5189–5196. doi: 10.1128/mcb.12.11.5189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lobo S. M., Lister J., Sullivan M. L., Hernandez N. The cloned RNA polymerase II transcription factor IID selects RNA polymerase III to transcribe the human U6 gene in vitro. Genes Dev. 1991 Aug;5(8):1477–1489. doi: 10.1101/gad.5.8.1477. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Meisterernst M., Roeder R. G. Family of proteins that interact with TFIID and regulate promoter activity. Cell. 1991 Nov 1;67(3):557–567. doi: 10.1016/0092-8674(91)90530-c. [DOI] [PubMed] [Google Scholar]
  16. Ranish J. A., Hahn S. The yeast general transcription factor TFIIA is composed of two polypeptide subunits. J Biol Chem. 1991 Oct 15;266(29):19320–19327. [PubMed] [Google Scholar]
  17. Ranish J. A., Lane W. S., Hahn S. Isolation of two genes that encode subunits of the yeast transcription factor IIA. Science. 1992 Feb 28;255(5048):1127–1129. doi: 10.1126/science.1546313. [DOI] [PubMed] [Google Scholar]
  18. Reinberg D., Horikoshi M., Roeder R. G. 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] [Google Scholar]
  19. Samuels M., Fire A., Sharp P. A. Separation and characterization of factors mediating accurate transcription by RNA polymerase II. J Biol Chem. 1982 Dec 10;257(23):14419–14427. [PubMed] [Google Scholar]
  20. Samuels M., Sharp P. A. Purification and characterization of a specific RNA polymerase II transcription factor. J Biol Chem. 1986 Feb 15;261(5):2003–2013. [PubMed] [Google Scholar]
  21. Sawadogo M., Sentenac A. RNA polymerase B (II) and general transcription factors. Annu Rev Biochem. 1990;59:711–754. doi: 10.1146/annurev.bi.59.070190.003431. [DOI] [PubMed] [Google Scholar]
  22. Schneider H. R., Waldschmidt R., Jahn D., Seifart K. H. Purification of human transcription factor IIIC and its binding to the gene for ribosomal 5S RNA. Nucleic Acids Res. 1989 Jul 11;17(13):5003–5016. doi: 10.1093/nar/17.13.5003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schneider H. R., Waldschmidt R., Seifart K. H. Human transcription factor IIIC contains a polypeptide of 55 kDa specifically binding to Pol III genes. Nucleic Acids Res. 1990 Aug 25;18(16):4743–4750. doi: 10.1093/nar/18.16.4743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schultz M. C., Reeder R. H., Hahn S. Variants of the TATA-binding protein can distinguish subsets of RNA polymerase I, II, and III promoters. Cell. 1992 May 15;69(4):697–702. doi: 10.1016/0092-8674(92)90233-3. [DOI] [PubMed] [Google Scholar]
  25. Segall J., Matsui T., Roeder R. G. Multiple factors are required for the accurate transcription of purified genes by RNA polymerase III. J Biol Chem. 1980 Dec 25;255(24):11986–11991. [PubMed] [Google Scholar]
  26. Seifart K. H., Wang L., Waldschmidt R., Jahn D., Wingender E. Purification of human transcription factor IIIA and its interaction with a chemically synthesized gene encoding human 5 S rRNA. J Biol Chem. 1989 Jan 25;264(3):1702–1709. [PubMed] [Google Scholar]
  27. Simmen K. A., Bernués J., Lewis J. D., Mattaj I. W. Cofractionation of the TATA-binding protein with the RNA polymerase III transcription factor TFIIIB. Nucleic Acids Res. 1992 Nov 25;20(22):5889–5898. doi: 10.1093/nar/20.22.5889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Simmen K. A., Bernués J., Parry H. D., Stunnenberg H. G., Berkenstam A., Cavallini B., Egly J. M., Mattaj I. W. TFIID is required for in vitro transcription of the human U6 gene by RNA polymerase III. EMBO J. 1991 Jul;10(7):1853–1862. doi: 10.1002/j.1460-2075.1991.tb07711.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Simmen K. A., Waldschmidt R., Bernués J., Parry H. D., Seifart K. H., Mattaj I. W. Proximal sequence element factor binding and species specificity in vertebrate U6 snRNA promoters. J Mol Biol. 1992 Feb 20;223(4):873–884. doi: 10.1016/0022-2836(92)90249-j. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Usuda Y., Kubota A., Berk A. J., Handa H. Affinity purification of transcription factor IIA from HeLa cell nuclear extracts. EMBO J. 1991 Aug;10(8):2305–2310. doi: 10.1002/j.1460-2075.1991.tb07767.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Waldschmidt R., Jahn D., Seifart K. H. Purification of transcription factor IIIB from HeLa cells. J Biol Chem. 1988 Sep 15;263(26):13350–13356. [PubMed] [Google Scholar]
  33. Waldschmidt R., Seifart K. H. TFIIA is required for in vitro transcription of mammalian U6 genes by RNA polymerase III. J Biol Chem. 1992 Aug 15;267(23):16359–16364. [PubMed] [Google Scholar]
  34. Waldschmidt R., Wanandi I., Seifart K. H. Identification of transcription factors required for the expression of mammalian U6 genes in vitro. EMBO J. 1991 Sep;10(9):2595–2603. doi: 10.1002/j.1460-2075.1991.tb07801.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. White R. J., Jackson S. P. Mechanism of TATA-binding protein recruitment to a TATA-less class III promoter. Cell. 1992 Dec 11;71(6):1041–1053. doi: 10.1016/0092-8674(92)90398-v. [DOI] [PubMed] [Google Scholar]
  36. White R. J., Jackson S. P., Rigby P. W. A role for the TATA-box-binding protein component of the transcription factor IID complex as a general RNA polymerase III transcription factor. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1949–1953. doi: 10.1073/pnas.89.5.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wingender E., Frank R., Blöcker H., Wang L. R., Jahn D., Seifart K. H. Complete synthesis and transcription in vitro of a gene coding for human ribosomal 5S RNA. Gene. 1988 Apr 15;64(1):77–85. doi: 10.1016/0378-1119(88)90482-9. [DOI] [PubMed] [Google Scholar]

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