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. 1996 Jun;16(6):2637–2646. doi: 10.1128/mcb.16.6.2637

Developmental silencing of the embryonic zeta-globin gene: concerted action of the promoter and the 3'-flanking region combined with stage-specific silencing by the transcribed segment.

S A Liebhaber 1, Z Wang 1, F E Cash 1, B Monks 1, J E Russell 1
PMCID: PMC231254  PMID: 8649371

Abstract

Globin gene switching is a well-described model of eucaryotic developmental control. In the case of the human alpha-globin gene cluster, migration of erythropoietic activity from the embryonic yolk sac to the fetal liver is parallaled by the zeta-globin gene silencing and enhanced expression of the alpha-globin genes. To map critical cis determinants of this switch, the human zeta-globin gene, the alpha-globin gene, and chimeric recombinants were introduced into the mouse genome. Consistent with previous studies, expression of the individual alpha- and zeta-globin transgenes was found to be developmentally appropriate. Contrary to current models, however, the alpha- and zeta-globin gene promoters were not sufficient to establish this control. Instead, full silencing of the zeta-globin gene required the combined activities of this promoter, transcribed region, and 3'-flanking sequences. Individually, the silencing activities of the zeta-globin gene promoter and 3'-flanking region were minimal but increased markedly when both regions were present. The zeta-globin transcribed region appeared to contribute to gene silencing by a mechanism specifically activated in definitive erythroblasts in the fetal liver. These data demonstrate that a complex set of controls, requiring at least three determinants and involving at least two independent mechanisms, is necessary for full developmental silencing of the human zeta-globin gene.

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Selected References

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  1. Albitar M., Care A., Peschle C., Liebhaber S. A. Developmental switching of messenger RNA expression from the human alpha-globin cluster: fetal/adult pattern of theta-globin gene expression. Blood. 1992 Sep 15;80(6):1586–1591. [PubMed] [Google Scholar]
  2. Albitar M., Cash F. E., Peschle C., Liebhaber S. A. Developmental switch in the relative expression of the alpha 1- and alpha 2-globin genes in humans and in transgenic mice. Blood. 1992 May 1;79(9):2471–2474. [PubMed] [Google Scholar]
  3. Albitar M., Katsumata M., Liebhaber S. A. Human alpha-globin genes demonstrate autonomous developmental regulation in transgenic mice. Mol Cell Biol. 1991 Jul;11(7):3786–3794. doi: 10.1128/mcb.11.7.3786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dillon N., Grosveld F. Transcriptional regulation of multigene loci: multilevel control. Trends Genet. 1993 Apr;9(4):134–137. doi: 10.1016/0168-9525(93)90208-y. [DOI] [PubMed] [Google Scholar]
  5. Engel J. D. Developmental regulation of human beta-globin gene transcription: a switch of loyalties? Trends Genet. 1993 Sep;9(9):304–309. doi: 10.1016/0168-9525(93)90248-g. [DOI] [PubMed] [Google Scholar]
  6. Goodman M., Czelusniak J., Koop B. F., Tagle D. A., Slightom J. L. Globins: a case study in molecular phylogeny. Cold Spring Harb Symp Quant Biol. 1987;52:875–890. doi: 10.1101/sqb.1987.052.01.096. [DOI] [PubMed] [Google Scholar]
  7. HUEHNS E. R., DANCE N., BEAVEN G. H., HECHT F., MOTULSKY A. G. HUMAN EMBRYONIC HEMOGLOBINS. Cold Spring Harb Symp Quant Biol. 1964;29:327–331. doi: 10.1101/sqb.1964.029.01.035. [DOI] [PubMed] [Google Scholar]
  8. Hanscombe O., Vidal M., Kaeda J., Luzzatto L., Greaves D. R., Grosveld F. High-level, erythroid-specific expression of the human alpha-globin gene in transgenic mice and the production of human hemoglobin in murine erythrocytes. Genes Dev. 1989 Oct;3(10):1572–1581. doi: 10.1101/gad.3.10.1572. [DOI] [PubMed] [Google Scholar]
  9. Higgs D. R., Vickers M. A., Wilkie A. O., Pretorius I. M., Jarman A. P., Weatherall D. J. A review of the molecular genetics of the human alpha-globin gene cluster. Blood. 1989 Apr;73(5):1081–1104. [PubMed] [Google Scholar]
  10. Higgs D. R., Wood W. G., Jarman A. P., Sharpe J., Lida J., Pretorius I. M., Ayyub H. A major positive regulatory region located far upstream of the human alpha-globin gene locus. Genes Dev. 1990 Sep;4(9):1588–1601. doi: 10.1101/gad.4.9.1588. [DOI] [PubMed] [Google Scholar]
  11. Kiledjian M., Wang X., Liebhaber S. A. Identification of two KH domain proteins in the alpha-globin mRNP stability complex. EMBO J. 1995 Sep 1;14(17):4357–4364. doi: 10.1002/j.1460-2075.1995.tb00110.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lau Y. F., Kan Y. W. Versatile cosmid vectors for the isolation, expression, and rescue of gene sequences: studies with the human alpha-globin gene cluster. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5225–5229. doi: 10.1073/pnas.80.17.5225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lauer J., Shen C. K., Maniatis T. The chromosomal arrangement of human alpha-like globin genes: sequence homology and alpha-globin gene deletions. Cell. 1980 May;20(1):119–130. doi: 10.1016/0092-8674(80)90240-8. [DOI] [PubMed] [Google Scholar]
  14. Leder A., Kuo A., Shen M. M., Leder P. In situ hybridization reveals co-expression of embryonic and adult alpha globin genes in the earliest murine erythrocyte progenitors. Development. 1992 Dec;116(4):1041–1049. doi: 10.1242/dev.116.4.1041. [DOI] [PubMed] [Google Scholar]
  15. Leder A., Swan D., Ruddle F., D'Eustachio P., Leder P. Dispersion of alpha-like globin genes of the mouse to three different chromosomes. Nature. 1981 Sep 17;293(5829):196–200. doi: 10.1038/293196a0. [DOI] [PubMed] [Google Scholar]
  16. Leder A., Weir L., Leder P. Characterization, expression, and evolution of the mouse embryonic zeta-globin gene. Mol Cell Biol. 1985 May;5(5):1025–1033. doi: 10.1128/mcb.5.5.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ley T. J., Maloney K. A., Gordon J. I., Schwartz A. L. Globin gene expression in erythroid human fetal liver cells. J Clin Invest. 1989 Mar;83(3):1032–1038. doi: 10.1172/JCI113944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liebhaber S. A. Alpha thalassemia. Hemoglobin. 1989;13(7-8):685–731. doi: 10.3109/03630268908998845. [DOI] [PubMed] [Google Scholar]
  19. Liebhaber S. A., Cash F. E., Ballas S. K. Human alpha-globin gene expression. The dominant role of the alpha 2-locus in mRNA and protein synthesis. J Biol Chem. 1986 Nov 15;261(32):15327–15333. [PubMed] [Google Scholar]
  20. Liebhaber S. A., Goossens M., Kan Y. W. Homology and concerted evolution at the alpha 1 and alpha 2 loci of human alpha-globin. Nature. 1981 Mar 5;290(5801):26–29. doi: 10.1038/290026a0. [DOI] [PubMed] [Google Scholar]
  21. Liebhaber S. A., Kan Y. W. Differentiation of the mRNA transcripts originating from the alpha 1- and alpha 2-globin loci in normals and alpha-thalassemics. J Clin Invest. 1981 Aug;68(2):439–446. doi: 10.1172/JCI110273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Modell B., Bulyzhenkov V. Distribution and control of some genetic disorders. World Health Stat Q. 1988;41(3-4):209–218. [PubMed] [Google Scholar]
  23. Moore M. A., Metcalf D. Ontogeny of the haemopoietic system: yolk sac origin of in vivo and in vitro colony forming cells in the developing mouse embryo. Br J Haematol. 1970 Mar;18(3):279–296. doi: 10.1111/j.1365-2141.1970.tb01443.x. [DOI] [PubMed] [Google Scholar]
  24. Peschle C., Mavilio F., Carè A., Migliaccio G., Migliaccio A. R., Salvo G., Samoggia P., Petti S., Guerriero R., Marinucci M. Haemoglobin switching in human embryos: asynchrony of zeta----alpha and epsilon----gamma-globin switches in primitive and definite erythropoietic lineage. Nature. 1985 Jan 17;313(5999):235–238. doi: 10.1038/313235a0. [DOI] [PubMed] [Google Scholar]
  25. Pondel M. D., Proudfoot N. J., Whitelaw C., Whitelaw E. The developmental regulation of the human zeta-globin gene in transgenic mice employing beta-galactosidase as a reporter gene. Nucleic Acids Res. 1992 Nov 11;20(21):5655–5660. doi: 10.1093/nar/20.21.5655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pressley L., Higgs D. R., Clegg J. B., Weatherall D. J. Gene deletions in alpha thalassemia prove that the 5' zeta locus is functional. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3586–3589. doi: 10.1073/pnas.77.6.3586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Purohit K., Deisseroth A. DNAase I nuclease digestion studies of the human zeta globin gene before and after transfer from the K562 cell to the mouse erythroleukemia cell. Prog Clin Biol Res. 1987;251:295–302. [PubMed] [Google Scholar]
  28. Raich N., Clegg C. H., Grofti J., Roméo P. H., Stamatoyannopoulos G. GATA1 and YY1 are developmental repressors of the human epsilon-globin gene. EMBO J. 1995 Feb 15;14(4):801–809. doi: 10.1002/j.1460-2075.1995.tb07058.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Raich N., Enver T., Nakamoto B., Josephson B., Papayannopoulou T., Stamatoyannopoulos G. Autonomous developmental control of human embryonic globin gene switching in transgenic mice. Science. 1990 Nov 23;250(4984):1147–1149. doi: 10.1126/science.2251502. [DOI] [PubMed] [Google Scholar]
  30. Russell E. S. Hereditary anemias of the mouse: a review for geneticists. Adv Genet. 1979;20:357–459. [PubMed] [Google Scholar]
  31. Sabath D. E., Koehler K. M., Yang W. Q., Patton K., Stamatoyannopoulos G. Identification of a major positive regulatory element located 5' to the human zeta-globin gene. Blood. 1995 May 1;85(9):2587–2597. [PubMed] [Google Scholar]
  32. Sabath D. E., Spangler E. A., Rubin E. M., Stamatoyannopoulos G. Analysis of the human zeta-globin gene promoter in transgenic mice. Blood. 1993 Nov 1;82(9):2899–2905. [PubMed] [Google Scholar]
  33. Sharpe J. A., Wells D. J., Whitelaw E., Vyas P., Higgs D. R., Wood W. G. Analysis of the human alpha-globin gene cluster in transgenic mice. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11262–11266. doi: 10.1073/pnas.90.23.11262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shih D. M., Wall R. J., Shapiro S. G. Developmentally regulated and erythroid-specific expression of the human embryonic beta-globin gene in transgenic mice. Nucleic Acids Res. 1990 Sep 25;18(18):5465–5472. doi: 10.1093/nar/18.18.5465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Spangler E. A., Andrews K. A., Rubin E. M. Developmental regulation of the human zeta globin gene in transgenic mice. Nucleic Acids Res. 1990 Dec 11;18(23):7093–7097. doi: 10.1093/nar/18.23.7093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Talbot D., Collis P., Antoniou M., Vidal M., Grosveld F., Greaves D. R. A dominant control region from the human beta-globin locus conferring integration site-independent gene expression. Nature. 1989 Mar 23;338(6213):352–355. doi: 10.1038/338352a0. [DOI] [PubMed] [Google Scholar]
  37. Tang W., Luo H. Y., Albitar M., Patterson M., Eng B., Waye J. S., Liebhaber S. A., Higgs D. R., Chui D. H. Human embryonic zeta-globin chain expression in deletional alpha-thalassemias. Blood. 1992 Jul 15;80(2):517–522. [PubMed] [Google Scholar]
  38. Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vyas P., Sharpe J. A., Watt P., Higgs D. R., Wood W. G. Regulation of human embryonic globin genes zeta 2 and epsilon in stably transformed mouse erythroleukemia cells. Blood. 1992 Oct 1;80(7):1832–1837. [PubMed] [Google Scholar]
  40. Vyas P., Vickers M. A., Simmons D. L., Ayyub H., Craddock C. F., Higgs D. R. Cis-acting sequences regulating expression of the human alpha-globin cluster lie within constitutively open chromatin. Cell. 1992 May 29;69(5):781–793. doi: 10.1016/0092-8674(92)90290-s. [DOI] [PubMed] [Google Scholar]
  41. Wang X., Kiledjian M., Weiss I. M., Liebhaber S. A. Detection and characterization of a 3' untranslated region ribonucleoprotein complex associated with human alpha-globin mRNA stability. Mol Cell Biol. 1995 Mar;15(3):1769–1777. doi: 10.1128/mcb.15.3.1769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weiss I. M., Liebhaber S. A. Erythroid cell-specific determinants of alpha-globin mRNA stability. Mol Cell Biol. 1994 Dec;14(12):8123–8132. doi: 10.1128/mcb.14.12.8123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Weiss I. M., Liebhaber S. A. Erythroid cell-specific mRNA stability elements in the alpha 2-globin 3' nontranslated region. Mol Cell Biol. 1995 May;15(5):2457–2465. doi: 10.1128/mcb.15.5.2457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Whitelaw E., Lamb P., Hogben P., Proudfoot N. J. The globin switch at the level of mRNA in the developing mouse. Prog Clin Biol Res. 1989;316A:323–333. [PubMed] [Google Scholar]
  45. Wong P. M., Chung S. W., Eaves C. J., Chui D. H. Ontogeny of the mouse hemopoietic system. Prog Clin Biol Res. 1985;193:17–28. [PubMed] [Google Scholar]
  46. Yagi M., Gelinas R., Elder J. T., Peretz M., Papayannopoulou T., Stamatoyannopoulos G., Groudine M. Chromatin structure and developmental expression of the human alpha-globin cluster. Mol Cell Biol. 1986 Apr;6(4):1108–1116. doi: 10.1128/mcb.6.4.1108. [DOI] [PMC free article] [PubMed] [Google Scholar]

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