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. 1996 Jun;16(6):2848–2856. doi: 10.1128/mcb.16.6.2848

Identification and analysis of a functional human homolog of the SPT4 gene of Saccharomyces cerevisiae.

G A Hartzog 1, M A Basrai 1, S L Ricupero-Hovasse 1, P Hieter 1, F Winston 1
PMCID: PMC231277  PMID: 8649394

Abstract

Spt4p is a nonhistone protein of Saccharomyces cerevisiae that is believed to be required for normal chromatin structure and transcription. In this work we describe the isolation and analysis of a human gene, SUPT4H, that encodes a predicted protein 42% identical to Spt4p. When expressed in S. cerevisiae, SUPT4H complemented all spt4 mutant phenotypes. In human cells SUPT4H encodes a nuclear protein that is expressed in all tissues tested. In addition, hybridization analyses suggest that an SUPT4H-related gene is also present in mice. SUPT4H was localized to human chromosome 17 by PCR analysis of a human-rodent somatic cell hybrid panel. Thus, like other proteins that are components of or control the structure of chromatin, Spt4p appears to be conserved from S. cerevisiae to mammals.

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Selected References

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

  1. Adams A. E., Shen W., Lin C. S., Leavitt J., Matsudaira P. Isoform-specific complementation of the yeast sac6 null mutation by human fimbrin. Mol Cell Biol. 1995 Jan;15(1):69–75. doi: 10.1128/mcb.15.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Aruffo A., Seed B. Molecular cloning of a CD28 cDNA by a high-efficiency COS cell expression system. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8573–8577. doi: 10.1073/pnas.84.23.8573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Basrai M. A., Kingsbury J., Koshland D., Spencer F., Hieter P. Faithful chromosome transmission requires Spt4p, a putative regulator of chromatin structure in Saccharomyces cerevisiae. Mol Cell Biol. 1996 Jun;16(6):2838–2847. doi: 10.1128/mcb.16.6.2838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bassett D. E., Jr, Boguski M. S., Spencer F., Reeves R., Goebl M., Hieter P. Comparative genomics, genome cross-referencing and XREFdb. Trends Genet. 1995 Sep;11(9):372–373. doi: 10.1016/s0168-9525(00)89109-x. [DOI] [PubMed] [Google Scholar]
  6. Baudin A., Ozier-Kalogeropoulos O., Denouel A., Lacroute F., Cullin C. A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae. Nucleic Acids Res. 1993 Jul 11;21(14):3329–3330. doi: 10.1093/nar/21.14.3329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  8. Boguski M. S., Lowe T. M., Tolstoshev C. M. dbEST--database for "expressed sequence tags". Nat Genet. 1993 Aug;4(4):332–333. doi: 10.1038/ng0893-332. [DOI] [PubMed] [Google Scholar]
  9. Buluwela L., Forster A., Boehm T., Rabbitts T. H. A rapid procedure for colony screening using nylon filters. Nucleic Acids Res. 1989 Jan 11;17(1):452–452. doi: 10.1093/nar/17.1.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Carlson M., Laurent B. C. The SNF/SWI family of global transcriptional activators. Curr Opin Cell Biol. 1994 Jun;6(3):396–402. doi: 10.1016/0955-0674(94)90032-9. [DOI] [PubMed] [Google Scholar]
  11. Clark-Adams C. D., Norris D., Osley M. A., Fassler J. S., Winston F. Changes in histone gene dosage alter transcription in yeast. Genes Dev. 1988 Feb;2(2):150–159. doi: 10.1101/gad.2.2.150. [DOI] [PubMed] [Google Scholar]
  12. Dingwall A. K., Beek S. J., McCallum C. M., Tamkun J. W., Kalpana G. V., Goff S. P., Scott M. P. The Drosophila snr1 and brm proteins are related to yeast SWI/SNF proteins and are components of a large protein complex. Mol Biol Cell. 1995 Jul;6(7):777–791. doi: 10.1091/mbc.6.7.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Drwinga H. L., Toji L. H., Kim C. H., Greene A. E., Mulivor R. A. NIGMS human/rodent somatic cell hybrid mapping panels 1 and 2. Genomics. 1993 May;16(2):311–314. doi: 10.1006/geno.1993.1190. [DOI] [PubMed] [Google Scholar]
  14. Elble R. A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992 Jul;13(1):18–20. [PubMed] [Google Scholar]
  15. Gyuris J., Golemis E., Chertkov H., Brent R. Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell. 1993 Nov 19;75(4):791–803. doi: 10.1016/0092-8674(93)90498-f. [DOI] [PubMed] [Google Scholar]
  16. Heald R., McLoughlin M., McKeon F. Human wee1 maintains mitotic timing by protecting the nucleus from cytoplasmically activated Cdc2 kinase. Cell. 1993 Aug 13;74(3):463–474. doi: 10.1016/0092-8674(93)80048-j. [DOI] [PubMed] [Google Scholar]
  17. Hereford L., Fahrner K., Woolford J., Jr, Rosbash M., Kaback D. B. Isolation of yeast histone genes H2A and H2B. Cell. 1979 Dec;18(4):1261–1271. doi: 10.1016/0092-8674(79)90237-x. [DOI] [PubMed] [Google Scholar]
  18. Hirschhorn J. N., Brown S. A., Clark C. D., Winston F. Evidence that SNF2/SWI2 and SNF5 activate transcription in yeast by altering chromatin structure. Genes Dev. 1992 Dec;6(12A):2288–2298. doi: 10.1101/gad.6.12a.2288. [DOI] [PubMed] [Google Scholar]
  19. Imbalzano A. N., Kwon H., Green M. R., Kingston R. E. Facilitated binding of TATA-binding protein to nucleosomal DNA. Nature. 1994 Aug 11;370(6489):481–485. doi: 10.1038/370481a0. [DOI] [PubMed] [Google Scholar]
  20. Kalpana G. V., Marmon S., Wang W., Crabtree G. R., Goff S. P. Binding and stimulation of HIV-1 integrase by a human homolog of yeast transcription factor SNF5. Science. 1994 Dec 23;266(5193):2002–2006. doi: 10.1126/science.7801128. [DOI] [PubMed] [Google Scholar]
  21. Khazak V., Sadhale P. P., Woychik N. A., Brent R., Golemis E. A. Human RNA polymerase II subunit hsRPB7 functions in yeast and influences stress survival and cell morphology. Mol Biol Cell. 1995 Jul;6(7):759–775. doi: 10.1091/mbc.6.7.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kwon H., Imbalzano A. N., Khavari P. A., Kingston R. E., Green M. R. Nucleosome disruption and enhancement of activator binding by a human SW1/SNF complex. Nature. 1994 Aug 11;370(6489):477–481. doi: 10.1038/370477a0. [DOI] [PubMed] [Google Scholar]
  23. Malone E. A., Fassler J. S., Winston F. Molecular and genetic characterization of SPT4, a gene important for transcription initiation in Saccharomyces cerevisiae. Mol Gen Genet. 1993 Mar;237(3):449–459. doi: 10.1007/BF00279450. [DOI] [PubMed] [Google Scholar]
  24. Maricq A. V., Peterson A. S., Brake A. J., Myers R. M., Julius D. Primary structure and functional expression of the 5HT3 receptor, a serotonin-gated ion channel. Science. 1991 Oct 18;254(5030):432–437. doi: 10.1126/science.1718042. [DOI] [PubMed] [Google Scholar]
  25. McAlpine P. Genetic nomenclature guide. Human. Trends Genet. 1995 Mar;:39–42. [PubMed] [Google Scholar]
  26. McKune K., Moore P. A., Hull M. W., Woychik N. A. Six human RNA polymerase subunits functionally substitute for their yeast counterparts. Mol Cell Biol. 1995 Dec;15(12):6895–6900. doi: 10.1128/mcb.15.12.6895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McKune K., Woychik N. A. Functional substitution of an essential yeast RNA polymerase subunit by a highly conserved mammalian counterpart. Mol Cell Biol. 1994 Jun;14(6):4155–4159. doi: 10.1128/mcb.14.6.4155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Meeks-Wagner D., Hartwell L. H. Normal stoichiometry of histone dimer sets is necessary for high fidelity of mitotic chromosome transmission. Cell. 1986 Jan 17;44(1):43–52. doi: 10.1016/0092-8674(86)90483-6. [DOI] [PubMed] [Google Scholar]
  29. Neigeborn L., Rubin K., Carlson M. Suppressors of SNF2 mutations restore invertase derepression and cause temperature-sensitive lethality in yeast. Genetics. 1986 Apr;112(4):741–753. doi: 10.1093/genetics/112.4.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Neiman A. M., Stevenson B. J., Xu H. P., Sprague G. F., Jr, Herskowitz I., Wigler M., Marcus S. Functional homology of protein kinases required for sexual differentiation in Schizosaccharomyces pombe and Saccharomyces cerevisiae suggests a conserved signal transduction module in eukaryotic organisms. Mol Biol Cell. 1993 Jan;4(1):107–120. doi: 10.1091/mbc.4.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nishiwaki K., Sano T., Miwa J. emb-5, a gene required for the correct timing of gut precursor cell division during gastrulation in Caenorhabditis elegans, encodes a protein similar to the yeast nuclear protein SPT6. Mol Gen Genet. 1993 Jun;239(3):313–322. doi: 10.1007/BF00276929. [DOI] [PubMed] [Google Scholar]
  32. Osborne M. A., Silver P. A. Nucleocytoplasmic transport in the yeast Saccharomyces cerevisiae. Annu Rev Biochem. 1993;62:219–254. doi: 10.1146/annurev.bi.62.070193.001251. [DOI] [PubMed] [Google Scholar]
  33. Paranjape S. M., Kamakaka R. T., Kadonaga J. T. Role of chromatin structure in the regulation of transcription by RNA polymerase II. Annu Rev Biochem. 1994;63:265–297. doi: 10.1146/annurev.bi.63.070194.001405. [DOI] [PubMed] [Google Scholar]
  34. Peterson C. L., Tamkun J. W. The SWI-SNF complex: a chromatin remodeling machine? Trends Biochem Sci. 1995 Apr;20(4):143–146. doi: 10.1016/s0968-0004(00)88990-2. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Spencer F., Gerring S. L., Connelly C., Hieter P. Mitotic chromosome transmission fidelity mutants in Saccharomyces cerevisiae. Genetics. 1990 Feb;124(2):237–249. doi: 10.1093/genetics/124.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Swanson M. S., Carlson M., Winston F. SPT6, an essential gene that affects transcription in Saccharomyces cerevisiae, encodes a nuclear protein with an extremely acidic amino terminus. Mol Cell Biol. 1990 Sep;10(9):4935–4941. doi: 10.1128/mcb.10.9.4935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Swanson M. S., Malone E. A., Winston F. SPT5, an essential gene important for normal transcription in Saccharomyces cerevisiae, encodes an acidic nuclear protein with a carboxy-terminal repeat. Mol Cell Biol. 1991 Jun;11(6):3009–3019. doi: 10.1128/mcb.11.6.3009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Swanson M. S., Winston F. SPT4, SPT5 and SPT6 interactions: effects on transcription and viability in Saccharomyces cerevisiae. Genetics. 1992 Oct;132(2):325–336. doi: 10.1093/genetics/132.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Takeda J., Espinosa R., 3rd, Eng S., Le Beau M. M., Bell G. I. Chromosomal assignment and tissue distribution of novel expressed sequence tags from a human pancreatic islet cDNA library. Genomics. 1995 Sep 1;29(1):276–281. doi: 10.1006/geno.1995.1245. [DOI] [PubMed] [Google Scholar]
  42. Wilson I. A., Niman H. L., Houghten R. A., Cherenson A. R., Connolly M. L., Lerner R. A. The structure of an antigenic determinant in a protein. Cell. 1984 Jul;37(3):767–778. doi: 10.1016/0092-8674(84)90412-4. [DOI] [PubMed] [Google Scholar]
  43. Winston F., Carlson M. Yeast SNF/SWI transcriptional activators and the SPT/SIN chromatin connection. Trends Genet. 1992 Nov;8(11):387–391. doi: 10.1016/0168-9525(92)90300-s. [DOI] [PubMed] [Google Scholar]
  44. Winston F., Dollard C., Ricupero-Hovasse S. L. Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C. Yeast. 1995 Jan;11(1):53–55. doi: 10.1002/yea.320110107. [DOI] [PubMed] [Google Scholar]
  45. Xu G. F., Lin B., Tanaka K., Dunn D., Wood D., Gesteland R., White R., Weiss R., Tamanoi F. The catalytic domain of the neurofibromatosis type 1 gene product stimulates ras GTPase and complements ira mutants of S. cerevisiae. Cell. 1990 Nov 16;63(4):835–841. doi: 10.1016/0092-8674(90)90149-9. [DOI] [PubMed] [Google Scholar]

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