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. 1991 Mar 11;19(5):1073–1079. doi: 10.1093/nar/19.5.1073

Cloning, expression and characterization of the human transcription elongation factor, TFIIS.

O J Yoo 1, H S Yoon 1, K H Baek 1, C J Jeon 1, K Miyamoto 1, A Ueno 1, K Agarwal 1
PMCID: PMC333783  PMID: 1708494

Abstract

The cDNA for the human elongation factor, TFIIS, has been cloned and expressed in E. coli with the T7 expression system. This 280-amino acid TFIIS protein is shorter by 21 residues than that of the mouse. The missing 21 residues are located in the amino-terminal region, which is not thought to be required for transcriptional stimulation. Apart from this gap, human and mouse proteins reveal 96% overall identity and 98.5% sequence similarity if conservative substitutions are taken into account. The bacterially expressed human protein and the purified calf thymus proteins are indistinguishable in their ability to stimulate transcript elongation by purified RNA polymerase II. Estimation of the native molecular size of the human protein in solution indicates that it exists as a dimer.

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Selected References

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

  1. Ahn B. Y., Gershon P. D., Jones E. V., Moss B. Identification of rpo30, a vaccinia virus RNA polymerase gene with structural similarity to a eucaryotic transcription elongation factor. Mol Cell Biol. 1990 Oct;10(10):5433–5441. doi: 10.1128/mcb.10.10.5433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baek K. H., Sato K., Ito R., Agarwal K. RNA polymerase II transcription terminates at a specific DNA sequence in a HeLa cell-free reaction. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7623–7627. doi: 10.1073/pnas.83.20.7623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Corden J. L. Tails of RNA polymerase II. Trends Biochem Sci. 1990 Oct;15(10):383–387. doi: 10.1016/0968-0004(90)90236-5. [DOI] [PubMed] [Google Scholar]
  4. Davies C. J., Trgovcich J., Hutchison C. A., 3rd Homologue of TFIIS in yeast. Nature. 1990 May 24;345(6273):298–298. doi: 10.1038/345298a0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Johnson P. F., McKnight S. L. Eukaryotic transcriptional regulatory proteins. Annu Rev Biochem. 1989;58:799–839. doi: 10.1146/annurev.bi.58.070189.004055. [DOI] [PubMed] [Google Scholar]
  8. Kadesch T. R., Chamberlin M. J. Studies of in vitro transcription by calf thymus RNA polymerase II using a novel duplex DNA template. J Biol Chem. 1982 May 10;257(9):5286–5295. [PubMed] [Google Scholar]
  9. Kerppola T. K., Kane C. M. Intrinsic sites of transcription termination and pausing in the c-myc gene. Mol Cell Biol. 1988 Oct;8(10):4389–4394. doi: 10.1128/mcb.8.10.4389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Landschulz W. H., Johnson P. F., Adashi E. Y., Graves B. J., McKnight S. L. Isolation of a recombinant copy of the gene encoding C/EBP. Genes Dev. 1988 Jul;2(7):786–800. doi: 10.1101/gad.2.7.786. [DOI] [PubMed] [Google Scholar]
  11. Lewin B. Commitment and activation at pol II promoters: a tail of protein-protein interactions. Cell. 1990 Jun 29;61(7):1161–1164. doi: 10.1016/0092-8674(90)90675-5. [DOI] [PubMed] [Google Scholar]
  12. Marshall T. K., Guo H., Price D. H. Drosophila RNA polymerase II elongation factor DmS-II has homology to mouse S-II and sequence similarity to yeast PPR2. Nucleic Acids Res. 1990 Nov 11;18(21):6293–6298. doi: 10.1093/nar/18.21.6293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McCauley R., Racker E. Separation of two monoamine oxidases from bovine bran. Mol Cell Biochem. 1973 May 11;1(1):73–81. doi: 10.1007/BF01659940. [DOI] [PubMed] [Google Scholar]
  14. Natori S. Stimulatory proteins of RNA polymerase II from Ehrlich ascites tumor cells. Mol Cell Biochem. 1982 Aug 6;46(3):173–187. doi: 10.1007/BF00239666. [DOI] [PubMed] [Google Scholar]
  15. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1–78. [PubMed] [Google Scholar]
  16. Rappaport J., Cho K., Saltzman A., Prenger J., Golomb M., Weinmann R. Transcription elongation factor SII interacts with a domain of the large subunit of human RNA polymerase II. Mol Cell Biol. 1988 Aug;8(8):3136–3142. doi: 10.1128/mcb.8.8.3136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sawadogo M., Sentenac A., Fromageot P. Interaction of a new polypeptide with yeast RNA polymerase B. J Biol Chem. 1980 Jan 10;255(1):12–15. [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. Sekimizu K., Nakanishi Y., Mizuno D., Natori S. Purification and preparation of antibody to RNA polymerase II stimulatory factors from Ehrlich ascites tumor cells. Biochemistry. 1979 Apr 17;18(8):1582–1588. doi: 10.1021/bi00575a031. [DOI] [PubMed] [Google Scholar]
  23. SivaRaman L., Reines D., Kane C. M. Purified elongation factor SII is sufficient to promote read-through by purified RNA polymerase II at specific termination sites in the human histone H3.3 gene. J Biol Chem. 1990 Aug 25;265(24):14554–14560. [PubMed] [Google Scholar]
  24. Sluder A. E., Greenleaf A. L., Price D. H. Properties of a Drosophila RNA polymerase II elongation factor. J Biol Chem. 1989 May 25;264(15):8963–8969. [PubMed] [Google Scholar]
  25. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]

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