Hormone-induced Recruitment of Sp1 Mediates Estrogen Activation of the Rabbit Uteroglobin Gene in Endometrial Epithelium*

Steroid hormones activate gene expression by interaction of their receptors with hormone-responsive DNA elements and tissue-specific or ubiquitous factors. To monitor the molecular changes on the promoter of the rabbit uteroglobin gene during early pseudopregnancy in vivo, we have applied the genomic footprinting methodology to endometrial tissue. Estrogen induction results in the simultaneous occupancy of an estrogen-responsive element and an adjacent GC/GT box in the promoter. DNA binding assays demonstrate that the corresponding regulatory factors are the ligand-induced estrogen receptor and the ubiquitous transcription factor Sp1. Both factors functionally synergize in primary endometrial cells, showing that the GC/GT box is an essential part of a composite estrogen-responsive unit. However, the estrogen receptor and Sp1 do not bind cooperatively to their sites in vitro, suggesting that other mechanisms might be responsible for the hormone-dependent binding of Sp1 in vivo. Since hormone treatment leads to the appearance of a distinct DNase I-hypersensitive site over the promoter chromatin, an estrogen-induced change in the local chromatin structure could facilitate binding of Sp1in vivo.

[1]  R. S. Krishnan,et al.  "Blastokinin": Inducer and Regulator of Blastocyst Development in the Rabbit Uterus , 1967, Science.

[2]  H. M. Beier,et al.  Uteroglobin: a hormone-sensitive endometrial protein involved in blastocyst development. , 1968, Biochimica et biophysica acta.

[3]  M. Beato,et al.  RNA synthesis in rabbit endometrial nuclei. Hormonal regulation of transcription of the uteroglobin gene. , 1980, European journal of biochemistry.

[4]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[5]  I. Dawid,et al.  Transient expression of genes introduced into cultured cells of Drosophila. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[6]  G. Church,et al.  Genomic sequencing. , 1993, Methods in molecular biology.

[7]  G. Greene,et al.  Immunochemical studies of estrogen receptors. , 1984, Journal of steroid biochemistry.

[8]  K. Kok,et al.  Nuclease-hypersensitive sites in chromatin of the estrogen-inducible apoVLDL II gene of chicken. , 1985, Nucleic acids research.

[9]  B. Gloss,et al.  In vivo protein–DNA interactions in a glucocorticoid response element require the presence of the hormone , 1986, Nature.

[10]  G. Hager,et al.  Steroid-dependent interaction of transcription factors with the inducible promoter of mouse mammary tumor virus in vivo , 1987, Cell.

[11]  M. Beato,et al.  Partial overlapping of binding sequences for steroid hormone receptors and DNaseI hypersensitive sites in the rabbit uteroglobin gene region. , 1987, Nucleic acids research.

[12]  H. Okayama,et al.  High-efficiency transformation of mammalian cells by plasmid DNA. , 1987, Molecular and cellular biology.

[13]  G. Schütz,et al.  Synergistic action of the glucocorticoid receptor with transcription factors. , 1988, The EMBO journal.

[14]  W. Herr,et al.  The ubiquitous octamer-binding protein Oct-1 contains a POU domain with a homeo box subdomain. , 1988, Genes & development.

[15]  R. Schüle,et al.  Cooperativity of the glucocorticoid receptor and the CACCC-box binding factor , 1988, Nature.

[16]  R. Tjian,et al.  Analysis of Sp1 in vivo reveals mutiple transcriptional domains, including a novel glutamine-rich activation motif , 1988, Cell.

[17]  J. Wijnholds,et al.  Tissue‐specific and steroid‐dependent interaction of transcription factors with the oestrogen‐inducible apoVLDL II promoter in vivo. , 1988, The EMBO journal.

[18]  S. Nordeen,et al.  Luciferase reporter gene vectors for analysis of promoters and enhancers. , 1988, BioTechniques.

[19]  R. Schüle,et al.  Many transcription factors interact synergistically with steroid receptors. , 1988, Science.

[20]  M. Schauer,et al.  Binding of hormone accelerates the kinetics of glucocorticoid and progesterone receptor binding to DNA. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[21]  B. Wold,et al.  Genomic sequencing and methylation analysis by ligation mediated PCR. , 1989, Science.

[22]  P. Chambon,et al.  The cloned human oestrogen receptor contains a mutation which alters its hormone binding properties. , 1989, The EMBO journal.

[23]  S. J. Thurston,et al.  SV40 stimulates expression of the transacting factor Sp1 at the mRNA level. , 1990, Genes & development.

[24]  M. Beato,et al.  The uteroglobin promoter contains a noncanonical estrogen responsive element. , 1990, Molecular endocrinology.

[25]  Jan Wijnholds,et al.  Oestrogen facilitates the binding of ubiquitous and liver-enriched nuclear proteins to the apoVLDL II promoter in vivo , 1991, Nucleic Acids Res..

[26]  N. Andrews,et al.  A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. , 1991, Nucleic acids research.

[27]  G. Chalepakis,et al.  Functional interaction of hybrid response elements with wild-type and mutant steroid hormone receptors , 1991, Molecular and cellular biology.

[28]  A. Gazdar,et al.  Elements of the rabbit uteroglobin promoter mediating its transcription in epithelial cells from the endometrium and lung. , 1992, Gene expression.

[29]  M. Beato,et al.  Cloning by recognition site screening of two novel GT box binding proteins: a family of Sp1 related genes. , 1992, Nucleic acids research.

[30]  R. Tjian,et al.  Molecular cloning and functional analysis of Drosophila TAF110 reveal properties expected of coactivators , 1993, Cell.

[31]  Y Fujii-Kuriyama,et al.  cDNA cloning and transcriptional properties of a novel GC box-binding protein, BTEB2. , 1993, Nucleic acids research.

[32]  T. Zacharewski,et al.  ICI 164,384: a control for investigating estrogen responsive genes. , 1993, Nucleic acids research.

[33]  R. V. van Waardenburg,et al.  Cis-acting elements reinforcing the activity of the estrogen-response element in the very-low-density apolipoprotein II gene promoter. , 1994, European journal of biochemistry.

[34]  X. Jacq,et al.  Human TAFII30 is present in a distinct TFIID complex and is required for transcriptional activation by the estrogen receptor , 1994, Cell.

[35]  M. Beato,et al.  Sp1‐mediated transcriptional activation is repressed by Sp3. , 1994, The EMBO journal.

[36]  H. Stunnenberg,et al.  Improved method for PCR-mediated site-directed mutagenesis. , 1994, Nucleic acids research.

[37]  R. Tjian,et al.  A glutamine-rich hydrophobic patch in transcription factor Sp1 contacts the dTAFII110 component of the Drosophila TFIID complex and mediates transcriptional activation. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[38]  C C Adams,et al.  Binding of disparate transcriptional activators to nucleosomal DNA is inherently cooperative , 1995, Molecular and cellular biology.

[39]  J. Widom,et al.  Mechanism of protein access to specific DNA sequences in chromatin: a dynamic equilibrium model for gene regulation. , 1995, Journal of molecular biology.

[40]  Miguel Beato,et al.  Steroid hormone receptors: Many Actors in search of a plot , 1995, Cell.

[41]  Robert Tjian,et al.  Multiple TAFIIs Directing Synergistic Activation of Transcription , 1995, Science.

[42]  M. Beato,et al.  Hormone induces binding of receptors and transcription factors to a rearranged nucleosome on the MMTV promoter in vivo. , 1995, The EMBO journal.

[43]  M. Beato,et al.  Members of the Sp Transcription Factor Family Control Transcription from the Uteroglobin Promoter (*) , 1995, The Journal of Biological Chemistry.

[44]  J. Direnzo,et al.  p300 is a component of an estrogen receptor coactivator complex. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[45]  G. Felsenfeld,et al.  Chromatin structure and gene expression. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[46]  M. Beato,et al.  An inhibitor domain in Sp3 regulates its glutamine‐rich activation domains. , 1996, The EMBO journal.

[47]  M. Beato,et al.  Binding of NF1 to the MMTV promoter in nucleosomes: influence of rotational phasing, translational positioning and histone H1. , 1997, Nucleic acids research.