Abstract
The DNaseI hypersensitive site 2 (HS2) of human β- globin locus control region (LCR) is required for the high level expression of human β-globin genes. In the present study, a stage-specific protein factor (LPF-β) was identified in the nuclear extract prepared from mouse fetal liver at d 18 of gestation, which could bind to the HS2 region of human β-globin LCR. We also found that the shift band of LPF-β factor could be competed by human β-globin promoter. However, it couldn't be competed by human ε-globin promoter or by human A-γ-globin promoter. Furthermore, our data demonstrated that the binding-sequence of LPF-β factor is 5'CACACCCTA 3', which is located at the HS2 region of β-LCR (from −10845 to −10853 bp) and human β-globin promoter (from −92 to −84 bp). We speculated that these regions containing the CACCC box in both the human β-globin promoter and HS2 might function as stage selector elements in the regulation of human β-globin switching and the LPF-β factor might be a stage-specific protein factor involved in the regulation of human β-globin gene expression.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Collins FS, Weissman SM . The molecular genetics of human hemoglobin. Prog Acid Res Mol Biol 1984; 31:315–462.
Tuan D, Solomon W, Li Q, London I . The “β-like globin” gene domain in human erythroid cells. Proc Natl Acad Sci USA 1985; 82:6384–8.
Grosveld F, Blom van Assendelft G, Greaves D, Kollias G . Position-independent high level expression of the human β-globin gene in transgenic mice. Cell 1987; 51:975–85.
Forrester W, Takegawa S, Papayannopoulou T, Stamatoyannopoulos G, Groudine M . Evidence for a locus activator region. Nucl Acids Res 1987; 15:10159–77.
Kioussis D, Vanin E, deLange T, Flavell RA, Grosveld F . β-globin gene inactivation by DNA translocation in γ-thalassemia. Nature 1983; 306:662–6.
Wright S, deBoer E, Rosenthal A, Flavell RA, Grosveld F . DNA sequences required for regulated expression of the β-globin genes in murine erythroleukaemia cells. Phil Trans R Soc Loud B 1984; 307:271–82.
Taramelli R, Kioussis D, Vanin E, Bartram K, Groffen J, Hurst J, Grosveld F . Gamma delta beta-thalassemias 1 and 2 are the result of a 100 kbp deletion in the human β-globin cluster. Nucl Acids Res 1986; 14:7017–29.
Chada K, Magram J, Costantini F . An embryonic pattern of expression of a human fetal globin gene in transgenic mice. Nature 1986; 319: 685–9.
Magram J, Chada K, Costantini F . Developmental regulation of a cloned adult β-globin gene in transgenic mice. Nature 1985; 315: 338–40.
Dillon N, Grosveld F . Human γ-globin genes silenced independently of other genes in the β- globin locus. Nature 1991; 350:252–4.
Enver T, Raich N, Ebens A J, Papayannopoulou T, Costantini F, Stamatoyannopoulos G . Developmental regulation of human fetal-to-adult globin gene switching in transgenic mice. Nature 1990; 344:309–13.
Gorski K, Carneiro M, Schibler U . Tissue-specific in vitro transcription from the mouse albumin promoter. Cell 1986; 47:767–76.
Bradford MM . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-Dye binding. Anal Biochem 1976; 72:248–54.
Strauss F, Varshavsky A . A protein binds to a satellite DNA repeat at three specific sites that would be brought into mutual proximity by DNA folding in the nucleosome. Cell 1984; 37:889–901.
Berg PE, Williams DM, Qian RL, et al. A common protein binds to two silencers 5′ to the human β-globin gene. Nucl Acids Res 1989; 17: 8833–52.
Strouboulis J, Dillon N, Grosveld F . Developmental regulation of a complete 70-Kb human β-globin locus in transgenic mice. Genes & Dev 1992; 6:1857–64.
Orkin SH, Antonarakis SE, Kazazian HH . Base substitution at position-88 in a β-thalassemic globin gene: further evidence for the role of distal promoter element ACACCC. J Biol Chem 1984; 259:8679–81.
Orkin SH, Kazazian HH, Giardina PJV . Linkage of β-thalassemia mutations and β-globin gene polymorphisms with DNA polymorphism in human β-globin gene cluster. Nature 1982; 296:627–31.
Mastrangelo I, Courey A, Wall J, Jackson S, Hough V . DNA looping and Sp1 multimer links: a mechanism for transcriptional synergism and enhancement. Proc Natl Acad Sci USA 1991; 88:5670–4.
Su W, Jackson S, Tjian R, Echols H . DNA looping between sites for transcriptional activation: self association of DNA-bound Sp1. Genes & Dev 1991; 5:820–6.
Mantovani R, Malgaretti N, Ottolenghi S . An erythroid specific nuclear factor binding to the proximal CACCC box of the β-globin gene promoter. Nucl Acids Res 1988; 16:4299–313.
Miller I, Bieker JJ . A novel, erythroid cell-specific murine transcription factor that binds to the CACCC element and is related to the Kruppel family of nuclear proteins. Mol & Cell Bio 1993; 13:2776–86.
Feng WC, Southwood CM, Bieker JJ . Analyses of β-thalassemia mutant DNA interactions with Erythroid Kruppel-like Factor (EKLF), an erythroid cell-specific transcription factor. J Biochem 1994; 269:1493–500.
Caterina J J, Ciavatta D J, Townes TM . Multiple elements in human β-globin locus control region 5' HS 2 are involved in enhancer activity and position-independent, transgene expression. Nucl Acids Res 1994; 22:1006–11.
Author information
Authors and Affiliations
Additional information
*Project was supported by grants from Shanghai Joint Laboratory of Life Sciences, Academia Sinica, and the National Natural Sciences Foundation.
Rights and permissions
About this article
Cite this article
Sun, T., Chen, Y., Hu, Y. et al. A stage-specific protein factor binding to a CACCC motif in both human β-globin gene promoter and 5'-HS2 region. Cell Res 4, 135–143 (1994). https://doi.org/10.1038/cr.1994.14
Received:
Revised:
Accepted:
Issue date:
DOI: https://doi.org/10.1038/cr.1994.14
