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. 2001 Jul 1;357(Pt 1):97–105. doi: 10.1042/0264-6021:3570097

Transcriptional regulation of the human cystathionine beta-synthase -1b basal promoter: synergistic transactivation by transcription factors NF-Y and Sp1/Sp3.

Y Ge 1, M A Konrad 1, L H Matherly 1, J W Taub 1
PMCID: PMC1221932  PMID: 11415440

Abstract

Cystathionine beta-synthase (CBS) catalyses the condensation of serine and homocysteine to form cystathionine, an intermediate step in the synthesis of cysteine. Human CBS encodes five distinct 5' non-coding exons, the most frequent termed CBS -1a and CBS -1b, each transcribed from its own unique GC-rich TATA-less promoter. The minimal transcriptional region (-3792 to -3667) of the CBS -1b promoter was defined by 5'- and 3'-deletions, and transient transfections of reporter gene constructs in HepG2 cells, characterized by CBS transcription exclusively from the -1b promoter. Included in this 125 bp region are 3 GC-boxes (termed GC-a, GC-b and GC-c), an inverted CAAT-box and an E-box. By gel-shift and supershift assays, binding of specificity protein (Sp)1 and Sp3 to the GC-box elements, upstream stimulatory factor 1 (USF-1) to the E-box, and both nuclear factor (NF)-Y and an NF-1-like factor to the CAAT box could be demonstrated. By transient trans fections and reporter gene assays in HepG2 and Drosophila SL2 cells, a functional interplay was indicated between NF-Y binding to the CAAT-box, or between USF-1 binding to the E-box, and Sp1/Sp3 binding to the GC-box elements. In SL2 cells, NF-Y and Sp1/Sp3 were synergistic. Furthermore, both Sp1 and the long Sp3 isoform transactivated the CBS -1b minimal promoter; however, the short Sp3 isoforms were potent repressors. These results may explain the cell- or tissue-specific regulation of CBS transcription, and clarify the bases for alterations in CBS gene expression in human disease and Down's syndrome.

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Selected References

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  1. Bao L., Vlcek C., Paces V., Kraus J. P. Identification and tissue distribution of human cystathionine beta-synthase mRNA isoforms. Arch Biochem Biophys. 1998 Feb 1;350(1):95–103. doi: 10.1006/abbi.1997.0486. [DOI] [PubMed] [Google Scholar]
  2. Chadefaux B., Ceballos I., Hamet M., Coude M., Poissonnier M., Kamoun P., Allard D. Is absence of atheroma in Down syndrome due to decreased homocysteine levels? Lancet. 1988 Sep 24;2(8613):741–741. doi: 10.1016/s0140-6736(88)90211-5. [DOI] [PubMed] [Google Scholar]
  3. Chassé J. F., Paul V., Escañez R., Kamoun P., London J. Human cystathionine beta-synthase: gene organization and expression of different 5' alternative splicing. Mamm Genome. 1997 Dec;8(12):917–921. doi: 10.1007/s003359900611. [DOI] [PubMed] [Google Scholar]
  4. Courey A. J., Holtzman D. A., Jackson S. P., Tjian R. Synergistic activation by the glutamine-rich domains of human transcription factor Sp1. Cell. 1989 Dec 1;59(5):827–836. doi: 10.1016/0092-8674(89)90606-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Dooley K. A., Bennett M. K., Osborne T. F. A critical role for cAMP response element-binding protein (CREB) as a Co-activator in sterol-regulated transcription of 3-hydroxy-3-methylglutaryl coenzyme A synthase promoter. J Biol Chem. 1999 Feb 26;274(9):5285–5291. doi: 10.1074/jbc.274.9.5285. [DOI] [PubMed] [Google Scholar]
  7. Födinger M., Buchmayer H., Sunder-Plassmann G. Molecular genetics of homocysteine metabolism. Miner Electrolyte Metab. 1999 Jul-Dec;25(4-6):269–278. doi: 10.1159/000057459. [DOI] [PubMed] [Google Scholar]
  8. Inoue T., Kamiyama J., Sakai T. Sp1 and NF-Y synergistically mediate the effect of vitamin D(3) in the p27(Kip1) gene promoter that lacks vitamin D response elements. J Biol Chem. 1999 Nov 5;274(45):32309–32317. doi: 10.1074/jbc.274.45.32309. [DOI] [PubMed] [Google Scholar]
  9. Kim I. S., Sinha S., de Crombrugghe B., Maity S. N. Determination of functional domains in the C subunit of the CCAAT-binding factor (CBF) necessary for formation of a CBF-DNA complex: CBF-B interacts simultaneously with both the CBF-A and CBF-C subunits to form a heterotrimeric CBF molecule. Mol Cell Biol. 1996 Aug;16(8):4003–4013. doi: 10.1128/mcb.16.8.4003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kraus J. P., Oliveriusová J., Sokolová J., Kraus E., Vlcek C., de Franchis R., Maclean K. N., Bao L., Bukovsk, Patterson D. The human cystathionine beta-synthase (CBS) gene: complete sequence, alternative splicing, and polymorphisms. Genomics. 1998 Sep 15;52(3):312–324. doi: 10.1006/geno.1998.5437. [DOI] [PubMed] [Google Scholar]
  11. Lange B. J., Kobrinsky N., Barnard D. R., Arthur D. C., Buckley J. D., Howells W. B., Gold S., Sanders J., Neudorf S., Smith F. O. Distinctive demography, biology, and outcome of acute myeloid leukemia and myelodysplastic syndrome in children with Down syndrome: Children's Cancer Group Studies 2861 and 2891. Blood. 1998 Jan 15;91(2):608–615. [PubMed] [Google Scholar]
  12. Li N., Seetharam B. A 69-base pair fragment derived from human transcobalamin II promoter is sufficient for high bidirectional activity in the absence of a TATA box and an initiator element in transfected cells. Role of an E box in transcriptional activity. J Biol Chem. 1998 Oct 23;273(43):28170–28177. doi: 10.1074/jbc.273.43.28170. [DOI] [PubMed] [Google Scholar]
  13. Liang F., Schaufele F., Gardner D. G. Functional interaction of NF-Y and Sp1 is required for type a natriuretic peptide receptor gene transcription. J Biol Chem. 2001 Jan 12;276(2):1516–1522. doi: 10.1074/jbc.M006350200. [DOI] [PubMed] [Google Scholar]
  14. Maity S. N., Vuorio T., de Crombrugghe B. The B subunit of a rat heteromeric CCAAT-binding transcription factor shows a striking sequence identity with the yeast Hap2 transcription factor. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5378–5382. doi: 10.1073/pnas.87.14.5378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mantovani R. The molecular biology of the CCAAT-binding factor NF-Y. Gene. 1999 Oct 18;239(1):15–27. doi: 10.1016/s0378-1119(99)00368-6. [DOI] [PubMed] [Google Scholar]
  16. Murakami Y., Ikeda U., Shimada K., Kawakami K. Promoter of the Na,K-ATPase alpha3 subunit gene is composed of cis elements to which NF-Y and Sp1/Sp3 bind in rat cardiocytes. Biochim Biophys Acta. 1997 Jun 26;1352(3):311–324. doi: 10.1016/s0167-4781(97)00032-8. [DOI] [PubMed] [Google Scholar]
  17. Pan J., McEver R. P. Characterization of the promoter for the human P-selectin gene. J Biol Chem. 1993 Oct 25;268(30):22600–22608. [PubMed] [Google Scholar]
  18. Ravindranath Y., Abella E., Krischer J. P., Wiley J., Inoue S., Harris M., Chauvenet A., Alvarado C. S., Dubowy R., Ritchey A. K. Acute myeloid leukemia (AML) in Down's syndrome is highly responsive to chemotherapy: experience on Pediatric Oncology Group AML Study 8498. Blood. 1992 Nov 1;80(9):2210–2214. [PubMed] [Google Scholar]
  19. Ribeiro A., Pastier D., Kardassis D., Chambaz J., Cardot P. Cooperative binding of upstream stimulatory factor and hepatic nuclear factor 4 drives the transcription of the human apolipoprotein A-II gene. J Biol Chem. 1999 Jan 15;274(3):1216–1225. doi: 10.1074/jbc.274.3.1216. [DOI] [PubMed] [Google Scholar]
  20. Roder K., Wolf S. S., Beck K. F., Schweizer M. Cooperative binding of NF-Y and Sp1 at the DNase I-hypersensitive site, fatty acid synthase insulin-responsive element 1, located at -500 in the rat fatty acid synthase promoter. J Biol Chem. 1997 Aug 22;272(34):21616–21624. doi: 10.1074/jbc.272.34.21616. [DOI] [PubMed] [Google Scholar]
  21. Roder K., Wolf S. S., Larkin K. J., Schweizer M. Interaction between the two ubiquitously expressed transcription factors NF-Y and Sp1. Gene. 1999 Jun 24;234(1):61–69. doi: 10.1016/s0378-1119(99)00180-8. [DOI] [PubMed] [Google Scholar]
  22. Roy A. L., Du H., Gregor P. D., Novina C. D., Martinez E., Roeder R. G. Cloning of an inr- and E-box-binding protein, TFII-I, that interacts physically and functionally with USF1. EMBO J. 1997 Dec 1;16(23):7091–7104. doi: 10.1093/emboj/16.23.7091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sieweke M. H., Tekotte H., Jarosch U., Graf T. Cooperative interaction of ets-1 with USF-1 required for HIV-1 enhancer activity in T cells. EMBO J. 1998 Mar 16;17(6):1728–1739. doi: 10.1093/emboj/17.6.1728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sinha S., Maity S. N., Lu J., de Crombrugghe B. Recombinant rat CBF-C, the third subunit of CBF/NFY, allows formation of a protein-DNA complex with CBF-A and CBF-B and with yeast HAP2 and HAP3. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1624–1628. doi: 10.1073/pnas.92.5.1624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sugiura N., Takishima K. Regulation of the gene promoter for extracellular signal-regulated protein kinase 2 by transcription factors NF-Y and Sp3. Biochem J. 2000 Apr 1;347(Pt 1):155–161. [PMC free article] [PubMed] [Google Scholar]
  26. Suske G. The Sp-family of transcription factors. Gene. 1999 Oct 1;238(2):291–300. doi: 10.1016/s0378-1119(99)00357-1. [DOI] [PubMed] [Google Scholar]
  27. Taub J. W., Huang X., Ge Y., Dutcher J. A., Stout M. L., Mohammad R. M., Ravindranath Y., Matherly L. H. Cystathionine-beta-synthase cDNA transfection alters the sensitivity and metabolism of 1-beta-D-arabinofuranosylcytosine in CCRF-CEM leukemia cells in vitro and in vivo: a model of leukemia in Down syndrome. Cancer Res. 2000 Nov 15;60(22):6421–6426. [PubMed] [Google Scholar]
  28. Taub J. W., Huang X., Matherly L. H., Stout M. L., Buck S. A., Massey G. V., Becton D. L., Chang M. N., Weinstein H. J., Ravindranath Y. Expression of chromosome 21-localized genes in acute myeloid leukemia: differences between Down syndrome and non-Down syndrome blast cells and relationship to in vitro sensitivity to cytosine arabinoside and daunorubicin. Blood. 1999 Aug 15;94(4):1393–1400. [PubMed] [Google Scholar]
  29. Taub J. W., Matherly L. H., Stout M. L., Buck S. A., Gurney J. G., Ravindranath Y. Enhanced metabolism of 1-beta-D-arabinofuranosylcytosine in Down syndrome cells: a contributing factor to the superior event free survival of Down syndrome children with acute myeloid leukemia. Blood. 1996 Apr 15;87(8):3395–3403. [PubMed] [Google Scholar]
  30. Taub J. W., Ravindranath Y. Treatment of childhood acute myeloid leukaemia. Baillieres Clin Haematol. 1996 Mar;9(1):129–146. doi: 10.1016/s0950-3536(96)80040-0. [DOI] [PubMed] [Google Scholar]
  31. Taub J. W., Stout M. L., Buck S. A., Huang X., Vega R. A., Becton D. L., Ravindranath Y. Myeloblasts from Down syndrome children with acute myeloid leukemia have increased in vitro sensitivity to cytosine arabinoside and daunorubicin. Leukemia. 1997 Sep;11(9):1594–1595. doi: 10.1038/sj.leu.2400747. [DOI] [PubMed] [Google Scholar]
  32. Ueland P. M., Refsum H., Christensen B. Methotrexate sensitivity in Down's syndrome: a hypothesis. Cancer Chemother Pharmacol. 1990;25(5):384–386. doi: 10.1007/BF00686245. [DOI] [PubMed] [Google Scholar]
  33. Vuorio T., Maity S. N., de Crombrugghe B. Purification and molecular cloning of the "A" chain of a rat heteromeric CCAAT-binding protein. Sequence identity with the yeast HAP3 transcription factor. J Biol Chem. 1990 Dec 25;265(36):22480–22486. [PubMed] [Google Scholar]
  34. Wang W., Dong L., Saville B., Safe S. Transcriptional activation of E2F1 gene expression by 17beta-estradiol in MCF-7 cells is regulated by NF-Y-Sp1/estrogen receptor interactions. Mol Endocrinol. 1999 Aug;13(8):1373–1387. doi: 10.1210/mend.13.8.0323. [DOI] [PubMed] [Google Scholar]
  35. Wright K. L., Moore T. L., Vilen B. J., Brown A. M., Ting J. P. Major histocompatibility complex class II-associated invariant chain gene expression is up-regulated by cooperative interactions of Sp1 and NF-Y. J Biol Chem. 1995 Sep 8;270(36):20978–20986. doi: 10.1074/jbc.270.36.20978. [DOI] [PubMed] [Google Scholar]
  36. Yamada K., Tanaka T., Miyamoto K., Noguchi T. Sp family members and nuclear factor-Y cooperatively stimulate transcription from the rat pyruvate kinase M gene distal promoter region via their direct interactions. J Biol Chem. 2000 Jun 16;275(24):18129–18137. doi: 10.1074/jbc.M001543200. [DOI] [PubMed] [Google Scholar]
  37. Zhong Z. D., Hammani K., Bae W. S., DeClerck Y. A. NF-Y and Sp1 cooperate for the transcriptional activation and cAMP response of human tissue inhibitor of metalloproteinases-2. J Biol Chem. 2000 Jun 16;275(24):18602–18610. doi: 10.1074/jbc.M001389200. [DOI] [PubMed] [Google Scholar]

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