Developmental regulation of the human embryonic beta-like globin gene is mediated by synergistic interactions among multiple tissue- and stage-specific elements

The stage-specific regulation of mammalian embryonic globin genes has been an experimentally elusive problem, in part because of the developmentally early timing of their expression. We have carried out a systematic analysis of truncation and internal deletion mutations within the 5'-flanking region of the human embryonic beta-like globin gene (epsilon) in erythroid and nonerythroid cell lines. Within a 670-bp region upstream from the constitutive promoter are multiple positive and negative control elements. Of these, a positive regulatory element (epsilon-PRE II) which is active only in embryonic erythroid cells is of particular interest. Remarkably, although it is inactive on its own, in the presence of other sequences located further upstream, it confers tissue- and developmental stage-specific expression on a constitutive epsilon-globin or heterologous promoter. The activity of epsilon-PRE II is also modulated by another positive regulatory domain located further downstream to direct erythroid cell-specific, but little or no embryonic stage-specific, transcription. A nuclear factor highly enriched in embryonic erythroid cells binds specifically within a 19-bp region of epsilon-PRE II. Nuclei from adult erythroid cells also contain a factor that binds to this region but forms a complex of faster electrophoretic mobility. We speculate that interactions between epsilon-PRE II and other upstream control elements play an important role in the developmental regulation of the human embryonic beta-like globin gene.

[1]  W Miller,et al.  Positive and negative regulatory elements of the rabbit embryonic epsilon-globin gene revealed by an improved multiple alignment program and functional analysis. , 1993, DNA sequence : the journal of DNA sequencing and mapping.

[2]  P. Marks,et al.  Erythroid cell development in fetal mice: ultrastructural characteristics and hemoglobin synthesis. , 1967, Journal of molecular biology.

[3]  F. Grosveld,et al.  Each hypersensitive site of the human beta-globin locus control region confers a different developmental pattern of expression on the globin genes. , 1993, Genes & development.

[4]  W. C. Forrester,et al.  Molecular analysis of the human beta-globin locus activation region. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[5]  J. D. Engel,et al.  The beta-globin stage selector element factor is erythroid-specific promoter/enhancer binding protein NF-E4. , 1989, Genes & Development.

[6]  S. Jane,et al.  Identification of a stage selector element in the human gamma‐globin gene promoter that fosters preferential interaction with the 5′ HS2 enhancer when in competition with the beta‐promoter. , 1992, The EMBO journal.

[7]  A. Carè,et al.  Haemoglobin switching in human embryos: asynchrony of ζ → α and ε → γ-globin switches in primitive and definitive erythropoietic lineage , 1985, Nature.

[8]  B. Seed,et al.  A simple phase-extraction assay for chloramphenicol acyltransferase activity. , 1988, Gene.

[9]  G. Stamatoyannopoulos,et al.  Autonomous developmental control of human embryonic globin gene switching in transgenic mice. , 1990, Science.

[10]  A. Bailey,et al.  CACC box and enhancer response of the human embryonic ε globin promoter , 1993 .

[11]  C. Y. Yu,et al.  The CACC box upstream of human embryonic epsilon globin gene binds Sp1 and is a functional promoter element in vitro and in vivo. , 1991, The Journal of biological chemistry.

[12]  T. Maniatis,et al.  Molecular cloning and characterization of the human beta-like globin gene cluster. , 1980, Cell.

[13]  S. Orkin.  Globin gene regulation and switching: Circa 1990 , 1990, Cell.

[14]  J. Lis,et al.  A germline transformation analysis reveals flexibility in the organization of heat shock consensus elements. , 1987, Nucleic acids research.

[15]  M. M. Garner,et al.  A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system , 1981, Nucleic Acids Res..

[16]  D. Kioussis,et al.  Differential expression of human globin genes introduced in K562 cells. , 1985, The EMBO journal.

[17]  F. Grosveld,et al.  Two tissue-specific factors bind the erythroid promoter of the human porphobilinogen deaminase gene. , 1989, Nucleic acids research.

[18]  S. Henikoff.  Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. , 1984, Gene.

[19]  M. Goodman,et al.  Phylogenetic footprinting reveals unexpected complexity in trans factor binding upstream from the epsilon-globin gene. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[20]  A. Schechter,et al.  Binding of erythroid and non-erythroid nuclear proteins to the silencer of the human ε-globin-encoding gene , 1992 .

[21]  J. D. Engel,et al.  Developmental regulation of β-globin gene switching , 1988, Cell.

[22]  D. Tuan,et al.  The "beta-like-globin" gene domain in human erythroid cells. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[23]  W. Schaffner,et al.  Cell type‐specificity elements of the immunoglobulin heavy chain gene enhancer. , 1987, The EMBO journal.

[24]  S. Weintraub,et al.  Retinoblastoma protein switches the E2F site from positive to negative element , 1992, Nature.

[25]  A. Maniatis.  Regulated Expression ofHumanot-and1-Globin Genes inTransient Heterokaryons , 1991 .

[26]  A. Schechter,et al.  Identification of a transcriptional silencer in the 5'-flanking region of the human epsilon-globin gene. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[27]  S. Orkin,et al.  Regulated expression of globin chains and the erythroid transcription factor GATA-1 during erythropoiesis in the developing mouse , 1990, Molecular and cellular biology.

[28]  N. Proudfoot,et al.  Distinct negative regulation of the human embryonic globin genes zeta and epsilon. , 1993, Gene expression.

[29]  T. Maniatis,et al.  Rapid reprogramming of globin gene expression in transient heterokaryons , 1986, Cell.

[30]  A. van der Eb,et al.  A new technique for the assay of infectivity of human adenovirus 5 DNA. , 1973, Virology.

[31]  James T. Elder,et al.  A developmentally stable chromatin structure in the human beta-globin gene cluster. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[32]  M. Baron,et al.  Reversibility of the differentiated state in somatic cells. , 1993, Current opinion in cell biology.

[33]  G. Kollias,et al.  Position-independent, high-level expression of the human β-globin gene in transgenic mice , 1987, Cell.

[34]  R. Myers,et al.  A directly repeated sequence in the beta-globin promoter regulates transcription in murine erythroleukemia cells , 1990, Molecular and cellular biology.

[35]  H. Blau,et al.  Differentiation requires continuous active control. , 1992, Annual review of biochemistry.

[36]  B. Luckow,et al.  CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements , 1987, Nucleic Acids Res..