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. 1990 Aug 25;18(16):4843–4849. doi: 10.1093/nar/18.16.4843

A heteromeric transcription factor required for mammalian RNA polymerase II.

S Kitajima 1, Y Tanaka 1, T Kawaguchi 1, T Nagaoka 1, S M Weissman 1, Y Yasukochi 1
PMCID: PMC331960  PMID: 2395645

Abstract

A general transcription factor, FC, essential for specific initiation of in vitro transcription by mammalian RNA polymerase II was identified and a procedure developed to purify it to near homogeneity from HeLa cell nuclei. Purified FC is composed of two polypeptides of apparent molecular masses 80 kDa and 30 kDa, on SDS-PAGE, and has a native size of 280 kDa estimated by gel filtration column. Both polypeptides were shown to be essential for reconstituting in vitro transcription activity. Biochemical analysis showed that the 80 kDa and 30 kDa components were present in a 1:1 molar ratio. FC was also demonstrated to interact directly or indirectly with purified RNA polymerase II. Similarities between FC and transcription factors reported by others from human, rat or Drosophila cells are discussed.

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Selected References

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  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Burton Z. F., Killeen M., Sopta M., Ortolan L. G., Greenblatt J. RAP30/74: a general initiation factor that binds to RNA polymerase II. Mol Cell Biol. 1988 Apr;8(4):1602–1613. doi: 10.1128/mcb.8.4.1602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burton Z. F., Ortolan L. G., Greenblatt J. Proteins that bind to RNA polymerase II are required for accurate initiation of transcription at the adenovirus 2 major late promoter. EMBO J. 1986 Nov;5(11):2923–2930. doi: 10.1002/j.1460-2075.1986.tb04588.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carthew R. W., Chodosh L. A., Sharp P. A. An RNA polymerase II transcription factor binds to an upstream element in the adenovirus major late promoter. Cell. 1985 Dec;43(2 Pt 1):439–448. doi: 10.1016/0092-8674(85)90174-6. [DOI] [PubMed] [Google Scholar]
  6. Cohen R. B., Yang L., Thompson J. A., Safer B. Identification of a downstream sequence and binding protein that regulate adenovirus major late promoter transcription in vitro. J Biol Chem. 1988 Jul 25;263(21):10377–10385. [PubMed] [Google Scholar]
  7. Conaway J. W., Conaway R. C. A multisubunit transcription factor essential for accurate initiation by RNA polymerase II. J Biol Chem. 1989 Feb 5;264(4):2357–2362. [PubMed] [Google Scholar]
  8. Davison B. L., Egly J. M., Mulvihill E. R., Chambon P. Formation of stable preinitiation complexes between eukaryotic class B transcription factors and promoter sequences. Nature. 1983 Feb 24;301(5902):680–686. doi: 10.1038/301680a0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Dignam J. D., Martin P. L., Shastry B. S., Roeder R. G. Eukaryotic gene transcription with purified components. Methods Enzymol. 1983;101:582–598. doi: 10.1016/0076-6879(83)01039-3. [DOI] [PubMed] [Google Scholar]
  11. Dynan W. S., Tjian R. The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell. 1983 Nov;35(1):79–87. doi: 10.1016/0092-8674(83)90210-6. [DOI] [PubMed] [Google Scholar]
  12. Fire A., Samuels M., Sharp P. A. Interactions between RNA polymerase II, factors, and template leading to accurate transcription. J Biol Chem. 1984 Feb 25;259(4):2509–2516. [PubMed] [Google Scholar]
  13. Flores O., Ha I., Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II. Purification and subunit composition of transcription factor IIF. J Biol Chem. 1990 Apr 5;265(10):5629–5634. [PubMed] [Google Scholar]
  14. Flores O., Maldonado E., Burton Z., Greenblatt J., Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II. RNA polymerase II-associating protein 30 is an essential component of transcription factor IIF. J Biol Chem. 1988 Aug 5;263(22):10812–10816. [PubMed] [Google Scholar]
  15. Flores O., Maldonado E., Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II. Factors IIE and IIF independently interact with RNA polymerase II. J Biol Chem. 1989 May 25;264(15):8913–8921. [PubMed] [Google Scholar]
  16. Hirashima S., Hirai H., Nakanishi Y., Natori S. Molecular cloning and characterization of cDNA for eukaryotic transcription factor S-II. J Biol Chem. 1988 Mar 15;263(8):3858–3863. [PubMed] [Google Scholar]
  17. Hodo H. G., 3rd, Blatti S. P. Purification using polyethylenimine precipitation and low molecular weight subunit analyses of calf thymus and wheat germ DNA-dependent RNA polymerase II. Biochemistry. 1977 May 31;16(11):2334–2343. doi: 10.1021/bi00630a005. [DOI] [PubMed] [Google Scholar]
  18. Horikoshi M., Sekimizu K., Natori S. Analysis of the stimulatory factor of RNA polymerase II in the initiation and elongation complex. J Biol Chem. 1984 Jan 10;259(1):608–611. [PubMed] [Google Scholar]
  19. Kitajima S., Kawaguchi T., Yasukochi Y., Weissman S. M. Resolution and partial characterization of factors required for in vitro transcription by mammalian RNA polymerase II. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6106–6110. doi: 10.1073/pnas.86.16.6106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Luse D. S., Roeder R. G. Accurate transcription initiation on a purified mouse beta-globin DNA fragment in a cell-free system. Cell. 1980 Jul;20(3):691–699. doi: 10.1016/0092-8674(80)90315-3. [DOI] [PubMed] [Google Scholar]
  22. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mansour S. L., Grodzicker T., Tjian R. Downstream sequences affect transcription initiation from the adenovirus major late promoter. Mol Cell Biol. 1986 Jul;6(7):2684–2694. doi: 10.1128/mcb.6.7.2684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  25. Matsui T., Segall J., Weil P. A., Roeder R. G. Multiple factors required for accurate initiation of transcription by purified RNA polymerase II. J Biol Chem. 1980 Dec 25;255(24):11992–11996. [PubMed] [Google Scholar]
  26. Miyamoto N. G., Moncollin V., Wintzerith M., Hen R., Egly J. M., Chambon P. Stimulation of in vitro transcription by the upstream element of the adenovirus-2 major late promoter involves a specific factor. Nucleic Acids Res. 1984 Dec 11;12(23):8779–8799. doi: 10.1093/nar/12.23.8779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moncollin V., Miyamoto N. G., Zheng X. M., Egly J. M. Purification of a factor specific for the upstream element of the adenovirus-2 major late promoter. EMBO J. 1986 Oct;5(10):2577–2584. doi: 10.1002/j.1460-2075.1986.tb04537.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nakajima N., Horikoshi M., Roeder R. G. Factors involved in specific transcription by mammalian RNA polymerase II: purification, genetic specificity, and TATA box-promoter interactions of TFIID. Mol Cell Biol. 1988 Oct;8(10):4028–4040. doi: 10.1128/mcb.8.10.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Price D. H., Sluder A. E., Greenleaf A. L. Dynamic interaction between a Drosophila transcription factor and RNA polymerase II. Mol Cell Biol. 1989 Apr;9(4):1465–1475. doi: 10.1128/mcb.9.4.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rappaport J., Reinberg D., Zandomeni R., Weinmann R. Purification and functional characterization of transcription factor SII from calf thymus. Role in RNA polymerase II elongation. J Biol Chem. 1987 Apr 15;262(11):5227–5232. [PubMed] [Google Scholar]
  31. 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]
  32. Reinberg D., Roeder R. G. 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] [Google Scholar]
  33. Reinberg D., Roeder R. G. Factors involved in specific transcription by mammalian RNA polymerase II. Transcription factor IIS stimulates elongation of RNA chains. J Biol Chem. 1987 Mar 5;262(7):3331–3337. [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. Sawadogo M., Roeder R. G. 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. doi: 10.1016/0092-8674(85)90021-2. [DOI] [PubMed] [Google Scholar]
  37. Sekimizu K., Kobayashi N., Mizuno D., Natori S. Purification of a factor from Ehrlich ascites tumor cells specifically stimulating RNA polymerase II. Biochemistry. 1976 Nov 16;15(23):5064–5070. doi: 10.1021/bi00668a018. [DOI] [PubMed] [Google Scholar]
  38. Shi X. P., Lee R., Weinmann R. Protein factor(s) binding independently to two different regions of the adenovirus 2 major late promoter. Nucleic Acids Res. 1986 May 12;14(9):3729–3744. doi: 10.1093/nar/14.9.3729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sopta M., Burton Z. F., Greenblatt J. Structure and associated DNA-helicase activity of a general transcription initiation factor that binds to RNA polymerase II. Nature. 1989 Oct 5;341(6241):410–414. doi: 10.1038/341410a0. [DOI] [PubMed] [Google Scholar]
  40. Sopta M., Carthew R. W., Greenblatt J. Isolation of three proteins that bind to mammalian RNA polymerase II. J Biol Chem. 1985 Aug 25;260(18):10353–10360. [PubMed] [Google Scholar]
  41. Switzer R. C., 3rd, Merril C. R., Shifrin S. A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal Biochem. 1979 Sep 15;98(1):231–237. doi: 10.1016/0003-2697(79)90732-2. [DOI] [PubMed] [Google Scholar]
  42. Tsai S. Y., Tsai M. J., Kops L. E., Minghetti P. P., O'Malley B. W. Transcription factors from oviduct and HeLa cells are similar. J Biol Chem. 1981 Dec 25;256(24):13055–13059. [PubMed] [Google Scholar]
  43. Wasylyk B., Chambon P. A T to A base substitution and small deletions in the conalbumin TATA box drastically decrease specific in vitro transcription. Nucleic Acids Res. 1981 Apr 24;9(8):1813–1824. doi: 10.1093/nar/9.8.1813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weil P. A., Luse D. S., Segall J., Roeder R. G. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979 Oct;18(2):469–484. doi: 10.1016/0092-8674(79)90065-5. [DOI] [PubMed] [Google Scholar]

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