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. 1998 Aug 1;26(15):3480–3485. doi: 10.1093/nar/26.15.3480

Structure and function of the human Werner syndrome gene promoter: evidence for transcriptional modulation.

L Wang 1, K E Hunt 1, G M Martin 1, J Oshima 1
PMCID: PMC147734  PMID: 9671808

Abstract

The Werner syndrome (WS) is an autosomal recessive segmental progeroid syndrome caused by mutations in a novel member ( WRN ) of the RecQ family of helicases. Somatic WS cells are hypermutable and have elongated S phases, suggesting possible defects in DNA replication and/or repair. As an initial approach to the investigation of how this locus might be responsive to DNA damage, we determined the structure of the human WRN promoter. The WRN promoter region has two transcription initiation sites and exhibits several features characteristic of so-called constitutive promoters, including the absence of TATA and CAAT boxes. A luciferase reporter assay revealed that the upstream promoter was used 2-10-fold less frequently than the downstream promoter, the variation being a function of cell type. The activity of the WRN promoter was dramatically reduced in cells from WS patients. The reduction of activity was not seen in three other promoters tested, including one TATA-less promoter and one TATA-containing promoter. This is consistent with the presence of a positive regulatory mechanism of WRN expression.

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Selected References

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  1. Azizkhan J. C., Jensen D. E., Pierce A. J., Wade M. Transcription from TATA-less promoters: dihydrofolate reductase as a model. Crit Rev Eukaryot Gene Expr. 1993;3(4):229–254. [PubMed] [Google Scholar]
  2. Bennett S. E., Umar A., Oshima J., Monnat R. J., Jr, Kunkel T. A. Mismatch repair in extracts of Werner syndrome cell lines. Cancer Res. 1997 Jul 15;57(14):2956–2960. [PubMed] [Google Scholar]
  3. Bird A. P. CpG-rich islands and the function of DNA methylation. Nature. 1986 May 15;321(6067):209–213. doi: 10.1038/321209a0. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Dynan W. S., Sazer S., Tjian R., Schimke R. T. Transcription factor Sp1 recognizes a DNA sequence in the mouse dihydrofolate reductase promoter. Nature. 1986 Jan 16;319(6050):246–248. doi: 10.1038/319246a0. [DOI] [PubMed] [Google Scholar]
  6. Ellis N. A., Groden J., Ye T. Z., Straughen J., Lennon D. J., Ciocci S., Proytcheva M., German J. The Bloom's syndrome gene product is homologous to RecQ helicases. Cell. 1995 Nov 17;83(4):655–666. doi: 10.1016/0092-8674(95)90105-1. [DOI] [PubMed] [Google Scholar]
  7. Epstein C. J., Martin G. M., Schultz A. L., Motulsky A. G. Werner's syndrome a review of its symptomatology, natural history, pathologic features, genetics and relationship to the natural aging process. Medicine (Baltimore) 1966 May;45(3):177–221. doi: 10.1097/00005792-196605000-00001. [DOI] [PubMed] [Google Scholar]
  8. Fukuchi K., Martin G. M., Monnat R. J., Jr Mutator phenotype of Werner syndrome is characterized by extensive deletions. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5893–5897. doi: 10.1073/pnas.86.15.5893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fukuchi K., Tanaka K., Kumahara Y., Marumo K., Pride M. B., Martin G. M., Monnat R. J., Jr Increased frequency of 6-thioguanine-resistant peripheral blood lymphocytes in Werner syndrome patients. Hum Genet. 1990 Feb;84(3):249–252. doi: 10.1007/BF00200569. [DOI] [PubMed] [Google Scholar]
  10. German J. Bloom syndrome: a mendelian prototype of somatic mutational disease. Medicine (Baltimore) 1993 Nov;72(6):393–406. [PubMed] [Google Scholar]
  11. Goddard K. A., Yu C. E., Oshima J., Miki T., Nakura J., Piussan C., Martin G. M., Schellenberg G. D., Wijsman E. M. Toward localization of the Werner syndrome gene by linkage disequilibrium and ancestral haplotyping: lessons learned from analysis of 35 chromosome 8p11.1-21.1 markers. Am J Hum Genet. 1996 Jun;58(6):1286–1302. [PMC free article] [PubMed] [Google Scholar]
  12. Gorbalenya A. E., Koonin E. V., Donchenko A. P., Blinov V. M. Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. Nucleic Acids Res. 1989 Jun 26;17(12):4713–4730. doi: 10.1093/nar/17.12.4713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goto M., Rubenstein M., Weber J., Woods K., Drayna D. Genetic linkage of Werner's syndrome to five markers on chromosome 8. Nature. 1992 Feb 20;355(6362):735–738. doi: 10.1038/355735a0. [DOI] [PubMed] [Google Scholar]
  14. Gray M. D., Shen J. C., Kamath-Loeb A. S., Blank A., Sopher B. L., Martin G. M., Oshima J., Loeb L. A. The Werner syndrome protein is a DNA helicase. Nat Genet. 1997 Sep;17(1):100–103. doi: 10.1038/ng0997-100. [DOI] [PubMed] [Google Scholar]
  15. Guzder S. N., Sung P., Bailly V., Prakash L., Prakash S. RAD25 is a DNA helicase required for DNA repair and RNA polymerase II transcription. Nature. 1994 Jun 16;369(6481):578–581. doi: 10.1038/369578a0. [DOI] [PubMed] [Google Scholar]
  16. Hanawalt P. C. Transcription-coupled repair and human disease. Science. 1994 Dec 23;266(5193):1957–1958. doi: 10.1126/science.7801121. [DOI] [PubMed] [Google Scholar]
  17. Hoehn H., Bryant E. M., Au K., Norwood T. H., Boman H., Martin G. M. Variegated translocation mosaicism in human skin fibroblast cultures. Cytogenet Cell Genet. 1975;15(5):282–298. doi: 10.1159/000130526. [DOI] [PubMed] [Google Scholar]
  18. Ishii S., Merlino G. T., Pastan I. Promoter region of the human Harvey ras proto-oncogene: similarity to the EGF receptor proto-oncogene promoter. Science. 1985 Dec 20;230(4732):1378–1381. doi: 10.1126/science.2999983. [DOI] [PubMed] [Google Scholar]
  19. Ishii S., Xu Y. H., Stratton R. H., Roe B. A., Merlino G. T., Pastan I. Characterization and sequence of the promoter region of the human epidermal growth factor receptor gene. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4920–4924. doi: 10.1073/pnas.82.15.4920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jacobson A., Peltz S. W. Interrelationships of the pathways of mRNA decay and translation in eukaryotic cells. Annu Rev Biochem. 1996;65:693–739. doi: 10.1146/annurev.bi.65.070196.003401. [DOI] [PubMed] [Google Scholar]
  21. Kadonaga J. T., Courey A. J., Ladika J., Tjian R. Distinct regions of Sp1 modulate DNA binding and transcriptional activation. Science. 1988 Dec 16;242(4885):1566–1570. doi: 10.1126/science.3059495. [DOI] [PubMed] [Google Scholar]
  22. Kageyama R., Merlino G. T., Pastan I. Nuclear factor ETF specifically stimulates transcription from promoters without a TATA box. J Biol Chem. 1989 Sep 15;264(26):15508–15514. [PubMed] [Google Scholar]
  23. Kageyama R., Pastan I. Molecular cloning and characterization of a human DNA binding factor that represses transcription. Cell. 1989 Dec 1;59(5):815–825. doi: 10.1016/0092-8674(89)90605-3. [DOI] [PubMed] [Google Scholar]
  24. Kill I. R., Faragher R. G., Lawrence K., Shall S. The expression of proliferation-dependent antigens during the lifespan of normal and progeroid human fibroblasts in culture. J Cell Sci. 1994 Feb;107(Pt 2):571–579. doi: 10.1242/jcs.107.2.571. [DOI] [PubMed] [Google Scholar]
  25. Kletzien R. F., Harris P. K., Foellmi L. A. Glucose-6-phosphate dehydrogenase: a "housekeeping" enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. FASEB J. 1994 Feb;8(2):174–181. doi: 10.1096/fasebj.8.2.8119488. [DOI] [PubMed] [Google Scholar]
  26. Liu J. M., Buchwald M., Walsh C. E., Young N. S. Fanconi anemia and novel strategies for therapy. Blood. 1994 Dec 15;84(12):3995–4007. [PubMed] [Google Scholar]
  27. Lohman T. M. Helicase-catalyzed DNA unwinding. J Biol Chem. 1993 Feb 5;268(4):2269–2272. [PubMed] [Google Scholar]
  28. Malkki M., Laakso M., Deeb S. S. The human hexokinase II gene promoter: functional characterization and detection of variants among patients with NIDDM. Diabetologia. 1997 Dec;40(12):1461–1469. doi: 10.1007/s001250050850. [DOI] [PubMed] [Google Scholar]
  29. Martin G. M. Genetic syndromes in man with potential relevance to the pathobiology of aging. Birth Defects Orig Artic Ser. 1978;14(1):5–39. [PubMed] [Google Scholar]
  30. Martin G. M., Sprague C. A., Epstein C. J. Replicative life-span of cultivated human cells. Effects of donor's age, tissue, and genotype. Lab Invest. 1970 Jul;23(1):86–92. [PubMed] [Google Scholar]
  31. Matsumoto T., Shimamoto A., Goto M., Furuichi Y. Impaired nuclear localization of defective DNA helicases in Werner's syndrome. Nat Genet. 1997 Aug;16(4):335–336. doi: 10.1038/ng0897-335. [DOI] [PubMed] [Google Scholar]
  32. McKeon C. Transcriptional regulation of the insulin receptor gene promoter. Adv Exp Med Biol. 1993;343:79–89. doi: 10.1007/978-1-4615-2988-0_9. [DOI] [PubMed] [Google Scholar]
  33. Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
  34. Moser M. J., Holley W. R., Chatterjee A., Mian I. S. The proofreading domain of Escherichia coli DNA polymerase I and other DNA and/or RNA exonuclease domains. Nucleic Acids Res. 1997 Dec 15;25(24):5110–5118. doi: 10.1093/nar/25.24.5110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mushegian A. R., Bassett D. E., Jr, Boguski M. S., Bork P., Koonin E. V. Positionally cloned human disease genes: patterns of evolutionary conservation and functional motifs. Proc Natl Acad Sci U S A. 1997 May 27;94(11):5831–5836. doi: 10.1073/pnas.94.11.5831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nakura J., Wijsman E. M., Miki T., Kamino K., Yu C. E., Oshima J., Fukuchi K., Weber J. L., Piussan C., Melaragno M. I. Homozygosity mapping of the Werner syndrome locus (WRN). Genomics. 1994 Oct;23(3):600–608. doi: 10.1006/geno.1994.1548. [DOI] [PubMed] [Google Scholar]
  37. Nance M. A., Berry S. A. Cockayne syndrome: review of 140 cases. Am J Med Genet. 1992 Jan 1;42(1):68–84. doi: 10.1002/ajmg.1320420115. [DOI] [PubMed] [Google Scholar]
  38. Oshima J., Yu C. E., Boehnke M., Weber J. L., Edelhoff S., Wagner M. J., Wells D. E., Wood S., Disteche C. M., Martin G. M. Integrated mapping analysis of the Werner syndrome region of chromosome 8. Genomics. 1994 Sep 1;23(1):100–113. doi: 10.1006/geno.1994.1464. [DOI] [PubMed] [Google Scholar]
  39. Oshima J., Yu C. E., Piussan C., Klein G., Jabkowski J., Balci S., Miki T., Nakura J., Ogihara T., Ells J. Homozygous and compound heterozygous mutations at the Werner syndrome locus. Hum Mol Genet. 1996 Dec;5(12):1909–1913. doi: 10.1093/hmg/5.12.1909. [DOI] [PubMed] [Google Scholar]
  40. Poot M., Hoehn H., Rünger T. M., Martin G. M. Impaired S-phase transit of Werner syndrome cells expressed in lymphoblastoid cell lines. Exp Cell Res. 1992 Oct;202(2):267–273. doi: 10.1016/0014-4827(92)90074-i. [DOI] [PubMed] [Google Scholar]
  41. Razin A., Kafri T. DNA methylation from embryo to adult. Prog Nucleic Acid Res Mol Biol. 1994;48:53–81. doi: 10.1016/s0079-6603(08)60853-3. [DOI] [PubMed] [Google Scholar]
  42. Rünger T. M., Bauer C., Dekant B., Möller K., Sobotta P., Czerny C., Poot M., Martin G. M. Hypermutable ligation of plasmid DNA ends in cells from patients with Werner syndrome. J Invest Dermatol. 1994 Jan;102(1):45–48. doi: 10.1111/1523-1747.ep12371730. [DOI] [PubMed] [Google Scholar]
  43. Salk D., Bryant E., Au K., Hoehn H., Martin G. M. Systematic growth studies, cocultivation, and cell hybridization studies of Werner syndrome cultured skin fibroblasts. Hum Genet. 1981;58(3):310–316. doi: 10.1007/BF00294930. [DOI] [PubMed] [Google Scholar]
  44. Salk D., Bryant E., Hoehn H., Johnston P., Martin G. M. Growth characteristics of Werner syndrome cells in vitro. Adv Exp Med Biol. 1985;190:305–311. doi: 10.1007/978-1-4684-7853-2_14. [DOI] [PubMed] [Google Scholar]
  45. Sassone-Corsi P., Lamph W. W., Kamps M., Verma I. M. fos-associated cellular p39 is related to nuclear transcription factor AP-1. Cell. 1988 Aug 12;54(4):553–560. doi: 10.1016/0092-8674(88)90077-3. [DOI] [PubMed] [Google Scholar]
  46. Sassone-Corsi P., Sisson J. C., Verma I. M. Transcriptional autoregulation of the proto-oncogene fos. Nature. 1988 Jul 28;334(6180):314–319. doi: 10.1038/334314a0. [DOI] [PubMed] [Google Scholar]
  47. Schellenberg G. D., Martin G. M., Wijsman E. M., Nakura J., Miki T., Ogihara T. Homozygosity mapping and Werner's syndrome. Lancet. 1992 Apr 18;339(8799):1002–1002. doi: 10.1016/0140-6736(92)91590-5. [DOI] [PubMed] [Google Scholar]
  48. Schmid S. R., Linder P. D-E-A-D protein family of putative RNA helicases. Mol Microbiol. 1992 Feb;6(3):283–291. doi: 10.1111/j.1365-2958.1992.tb01470.x. [DOI] [PubMed] [Google Scholar]
  49. Shiroishi T., Koide T., Yoshino M., Sagai T., Moriwaki K. Hotspots of homologous recombination in mouse meiosis. Adv Biophys. 1995;31:119–132. doi: 10.1016/0065-227x(95)99387-5. [DOI] [PubMed] [Google Scholar]
  50. Smith C. L., Hager G. L. Transcriptional regulation of mammalian genes in vivo. A tale of two templates. J Biol Chem. 1997 Oct 31;272(44):27493–27496. doi: 10.1074/jbc.272.44.27493. [DOI] [PubMed] [Google Scholar]
  51. Stern H., Hotta Y. The organization of DNA metabolism during the recombinational phase of meiosis with special reference to humans. Mol Cell Biochem. 1980 Feb 28;29(3):145–158. doi: 10.1007/BF00420286. [DOI] [PubMed] [Google Scholar]
  52. Sung P., Bailly V., Weber C., Thompson L. H., Prakash L., Prakash S. Human xeroderma pigmentosum group D gene encodes a DNA helicase. Nature. 1993 Oct 28;365(6449):852–855. doi: 10.1038/365852a0. [DOI] [PubMed] [Google Scholar]
  53. Suzuki N., Shimamoto A., Imamura O., Kuromitsu J., Kitao S., Goto M., Furuichi Y. DNA helicase activity in Werner's syndrome gene product synthesized in a baculovirus system. Nucleic Acids Res. 1997 Aug 1;25(15):2973–2978. doi: 10.1093/nar/25.15.2973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Troelstra C., van Gool A., de Wit J., Vermeulen W., Bootsma D., Hoeijmakers J. H. ERCC6, a member of a subfamily of putative helicases, is involved in Cockayne's syndrome and preferential repair of active genes. Cell. 1992 Dec 11;71(6):939–953. doi: 10.1016/0092-8674(92)90390-x. [DOI] [PubMed] [Google Scholar]
  55. Tuteja N., Tuteja R. DNA helicases: the long unwinding road. Nat Genet. 1996 May;13(1):11–12. doi: 10.1038/ng0596-11. [DOI] [PubMed] [Google Scholar]
  56. Umezu K., Nakayama K., Nakayama H. Escherichia coli RecQ protein is a DNA helicase. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5363–5367. doi: 10.1073/pnas.87.14.5363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Webb D. K., Evans M. K., Bohr V. A. DNA repair fine structure in Werner's syndrome cell lines. Exp Cell Res. 1996 May 1;224(2):272–278. doi: 10.1006/excr.1996.0137. [DOI] [PubMed] [Google Scholar]
  58. Weber C. A., Salazar E. P., Stewart S. A., Thompson L. H. ERCC2: cDNA cloning and molecular characterization of a human nucleotide excision repair gene with high homology to yeast RAD3. EMBO J. 1990 May;9(5):1437–1447. doi: 10.1002/j.1460-2075.1990.tb08260.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Weeda G., Ma L., van Ham R. C., Bootsma D., van der Eb A. J., Hoeijmakers J. H. Characterization of the mouse homolog of the XPBC/ERCC-3 gene implicated in xeroderma pigmentosum and Cockayne's syndrome. Carcinogenesis. 1991 Dec;12(12):2361–2368. doi: 10.1093/carcin/12.12.2361. [DOI] [PubMed] [Google Scholar]
  60. Weeda G., van Ham R. C., Vermeulen W., Bootsma D., van der Eb A. J., Hoeijmakers J. H. A presumed DNA helicase encoded by ERCC-3 is involved in the human repair disorders xeroderma pigmentosum and Cockayne's syndrome. Cell. 1990 Aug 24;62(4):777–791. doi: 10.1016/0092-8674(90)90122-u. [DOI] [PubMed] [Google Scholar]
  61. West S. C. The processing of recombination intermediates: mechanistic insights from studies of bacterial proteins. Cell. 1994 Jan 14;76(1):9–15. doi: 10.1016/0092-8674(94)90168-6. [DOI] [PubMed] [Google Scholar]
  62. Yamabe Y., Sugimoto M., Satoh M., Suzuki N., Sugawara M., Goto M., Furuichi Y. Down-regulation of the defective transcripts of the Werner's syndrome gene in the cells of patients. Biochem Biophys Res Commun. 1997 Jul 9;236(1):151–154. doi: 10.1006/bbrc.1997.6919. [DOI] [PubMed] [Google Scholar]
  63. Yang W. S., Nevin D. N., Peng R., Brunzell J. D., Deeb S. S. A mutation in the promoter of the lipoprotein lipase (LPL) gene in a patient with familial combined hyperlipidemia and low LPL activity. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4462–4466. doi: 10.1073/pnas.92.10.4462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Yu C. E., Oshima J., Fu Y. H., Wijsman E. M., Hisama F., Alisch R., Matthews S., Nakura J., Miki T., Ouais S. Positional cloning of the Werner's syndrome gene. Science. 1996 Apr 12;272(5259):258–262. doi: 10.1126/science.272.5259.258. [DOI] [PubMed] [Google Scholar]
  65. Yu C. E., Oshima J., Goddard K. A., Miki T., Nakura J., Ogihara T., Poot M., Hoehn H., Fraccaro M., Piussan C. Linkage disequilibrium and haplotype studies of chromosome 8p 11.1-21.1 markers and Werner syndrome. Am J Hum Genet. 1994 Aug;55(2):356–364. [PMC free article] [PubMed] [Google Scholar]
  66. Yu C. E., Oshima J., Wijsman E. M., Nakura J., Miki T., Piussan C., Matthews S., Fu Y. H., Mulligan J., Martin G. M. Mutations in the consensus helicase domains of the Werner syndrome gene. Werner's Syndrome Collaborative Group. Am J Hum Genet. 1997 Feb;60(2):330–341. [PMC free article] [PubMed] [Google Scholar]

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