Abstract
The -198 T----C mutation in the promoter of the A gamma-globin gene increases 20-30 fold the expression of this gene in adult erythroid cells of patients (Hereditary Persistence of Fetal Hemoglobin, HPFH). We show here that this mutation creates a strong binding site, resembling a CACCC box, for two ubiquitous nuclear proteins, one of which is Sp1. The mutated promoter is four to five-fold more efficient than a normal gamma-globin promoter in driving expression of a CAT reporter plasmid when transfected into erythroid cells. The overexpression of the mutant is abolished by the introduction of an additional mutation disrupting the new binding site. No overexpression of the mutant is observed in non-erythroid cells, indicating that the ubiquitous factors bound on the mutated sequence must cooperate with erythroid specific factors.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Briggs M. R., Kadonaga J. T., Bell S. P., Tjian R. Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science. 1986 Oct 3;234(4772):47–52. doi: 10.1126/science.3529394. [DOI] [PubMed] [Google Scholar]
- Catala F., deBoer E., Habets G., Grosveld F. Nuclear protein factors and erythroid transcription of the human A gamma-globin gene. Nucleic Acids Res. 1989 May 25;17(10):3811–3827. doi: 10.1093/nar/17.10.3811. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collins F. S., Stoeckert C. J., Jr, Serjeant G. R., Forget B. G., Weissman S. M. G gamma beta+ hereditary persistence of fetal hemoglobin: cosmid cloning and identification of a specific mutation 5' to the G gamma gene. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4894–4898. doi: 10.1073/pnas.81.15.4894. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davidson I., Xiao J. H., Rosales R., Staub A., Chambon P. The HeLa cell protein TEF-1 binds specifically and cooperatively to two SV40 enhancer motifs of unrelated sequence. Cell. 1988 Sep 23;54(7):931–942. doi: 10.1016/0092-8674(88)90108-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Emerson B. M., Nickol J. M., Fong T. C. Erythroid-specific activation and derepression of the chick beta-globin promoter in vitro. Cell. 1989 Jun 30;57(7):1189–1200. doi: 10.1016/0092-8674(89)90056-1. [DOI] [PubMed] [Google Scholar]
- Gelinas R., Bender M., Lotshaw C., Waber P., Kazazian H., Jr, Stamatoyannopoulos G. Chinese A gamma fetal hemoglobin: C to T substitution at position-196 of the A gamma gene promoter. Blood. 1986 Jun;67(6):1777–1779. [PubMed] [Google Scholar]
- Giglioni B., Casini C., Mantovani R., Merli S., Comi P., Ottolenghi S., Saglio G., Camaschella C., Mazza U. A molecular study of a family with Greek hereditary persistence of fetal hemoglobin and beta-thalassemia. EMBO J. 1984 Nov;3(11):2641–2645. doi: 10.1002/j.1460-2075.1984.tb02187.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giglioni B., Comi P., Ronchi A., Mantovani R., Ottolenghi S. The same nuclear proteins bind the proximal CACCC box of the human beta-globin promoter and a similar sequence in the enhancer. Biochem Biophys Res Commun. 1989 Oct 16;164(1):149–155. doi: 10.1016/0006-291x(89)91695-1. [DOI] [PubMed] [Google Scholar]
- Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hooft van Huijsduijnen R. A., Bollekens J., Dorn A., Benoist C., Mathis D. Properties of a CCAAT box-binding protein. Nucleic Acids Res. 1987 Sep 25;15(18):7265–7282. doi: 10.1093/nar/15.18.7265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones K. A., Kadonaga J. T., Luciw P. A., Tjian R. Activation of the AIDS retrovirus promoter by the cellular transcription factor, Sp1. Science. 1986 May 9;232(4751):755–759. doi: 10.1126/science.3008338. [DOI] [PubMed] [Google Scholar]
- Mantovani R., Malgaretti N., Giglioni B., Comi P., Cappellini N., Nicolis S., Ottolenghi S. A protein factor binding to an octamer motif in the gamma-globin promoter disappears upon induction of differentiation and hemoglobin synthesis in K562 cells. Nucleic Acids Res. 1987 Nov 25;15(22):9349–9364. doi: 10.1093/nar/15.22.9349. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mantovani R., Malgaretti N., Nicolis S., Giglioni B., Comi P., Cappellini N., Bertero M. T., Caligaris-Cappio F., Ottolenghi S. An erythroid specific nuclear factor binding to the proximal CACCC box of the beta-globin gene promoter. Nucleic Acids Res. 1988 May 25;16(10):4299–4313. doi: 10.1093/nar/16.10.4299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mantovani R., Malgaretti N., Nicolis S., Ronchi A., Giglioni B., Ottolenghi S. The effects of HPFH mutations in the human gamma-globin promoter on binding of ubiquitous and erythroid specific nuclear factors. Nucleic Acids Res. 1988 Aug 25;16(16):7783–7797. doi: 10.1093/nar/16.16.7783. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mantovani R., Superti-Furga G., Gilman J., Ottolenghi S. The deletion of the distal CCAAT box region of the A gamma-globin gene in black HPFH abolishes the binding of the erythroid specific protein NFE3 and of the CCAAT displacement protein. Nucleic Acids Res. 1989 Aug 25;17(16):6681–6691. doi: 10.1093/nar/17.16.6681. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martin D. I., Tsai S. F., Orkin S. H. Increased gamma-globin expression in a nondeletion HPFH mediated by an erythroid-specific DNA-binding factor. Nature. 1989 Mar 30;338(6214):435–438. doi: 10.1038/338435a0. [DOI] [PubMed] [Google Scholar]
- Nicolis S., Ronchi A., Malgaretti N., Mantovani R., Giglioni B., Ottolenghi S. Increased erythroid-specific expression of a mutated HPFH gamma-globin promoter requires the erythroid factor NFE-1. Nucleic Acids Res. 1989 Jul 25;17(14):5509–5516. doi: 10.1093/nar/17.14.5509. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schüle R., Muller M., Kaltschmidt C., Renkawitz R. Many transcription factors interact synergistically with steroid receptors. Science. 1988 Dec 9;242(4884):1418–1420. doi: 10.1126/science.3201230. [DOI] [PubMed] [Google Scholar]
- Seed B., Sheen J. Y. A simple phase-extraction assay for chloramphenicol acyltransferase activity. Gene. 1988 Jul 30;67(2):271–277. doi: 10.1016/0378-1119(88)90403-9. [DOI] [PubMed] [Google Scholar]
- Singh H., Sen R., Baltimore D., Sharp P. A. A nuclear factor that binds to a conserved sequence motif in transcriptional control elements of immunoglobulin genes. Nature. 1986 Jan 9;319(6049):154–158. doi: 10.1038/319154a0. [DOI] [PubMed] [Google Scholar]
- Superti-Furga G., Barberis A., Schaffner G., Busslinger M. The -117 mutation in Greek HPFH affects the binding of three nuclear factors to the CCAAT region of the gamma-globin gene. EMBO J. 1988 Oct;7(10):3099–3107. doi: 10.1002/j.1460-2075.1988.tb03176.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tate V. E., Wood W. G., Weatherall D. J. The British form of hereditary persistence of fetal hemoglobin results from a single base mutation adjacent to an S1 hypersensitive site 5' to the A gamma globin gene. Blood. 1986 Dec;68(6):1389–1393. [PubMed] [Google Scholar]
- Treisman R., Orkin S. H., Maniatis T. Specific transcription and RNA splicing defects in five cloned beta-thalassaemia genes. Nature. 1983 Apr 14;302(5909):591–596. doi: 10.1038/302591a0. [DOI] [PubMed] [Google Scholar]
- Wood W. G., MacRae I. A., Darbre P. D., Clegg J. B., Weatherall D. J. The British type of non-deletion HPFH: characterization of developmental changes in vivo and erythroid growth in vitro. Br J Haematol. 1982 Mar;50(3):401–414. doi: 10.1111/j.1365-2141.1982.tb01935.x. [DOI] [PubMed] [Google Scholar]
- Xiao J. H., Davidson I., Macchi M., Rosales R., Vigneron M., Staub A., Chambon P. In vitro binding of several cell-specific and ubiquitous nuclear proteins to the GT-I motif of the SV40 enhancer. Genes Dev. 1987 Oct;1(8):794–807. doi: 10.1101/gad.1.8.794. [DOI] [PubMed] [Google Scholar]
- Yang K. G., Stoming T. A., Fei Y. J., Liang S., Wong S. C., Masala B., Huang R. B., Wei Z. P., Huisman T. H. Identification of base substitutions in the promoter regions of the A gamma- and G gamma-globin genes in A gamma- (or G gamma-) beta+-HPFH heterozygotes using the DNA-amplification-synthetic oligonucleotide procedure. Blood. 1988 May;71(5):1414–1417. [PubMed] [Google Scholar]
- deBoer E., Antoniou M., Mignotte V., Wall L., Grosveld F. The human beta-globin promoter; nuclear protein factors and erythroid specific induction of transcription. EMBO J. 1988 Dec 20;7(13):4203–4212. doi: 10.1002/j.1460-2075.1988.tb03317.x. [DOI] [PMC free article] [PubMed] [Google Scholar]