Hormone‐induced nucleosome positioning in the MMTV promoter is reversible

The mouse mammary tumor virus (MMTV) promoter is induced by glucocorticoid hormone via the glucocorticoid receptor (GR). The hormone‐triggered effects on MMTV transcription and chromatin structure were studied in Xenopus oocytes. We previously showed that the nucleosomes organizing the MMTV promoter became translationally positioned upon hormone induction. A single GR‐binding site was necessary and sufficient for the chromatin events to occur, while transcription and basal promoter elements were dispensable. Here we show that addition of the hormone antagonists RU486 or RU43044 to the previously hormone‐induced MMTV promoter results in cessation of transcription and loss of chromatin remodeling and nucleosome positioning. In vivo footprinting demonstrated agonist‐ and RU486‐induced GR binding to its DNA response element (GRE), while the other antagonist, RU43044, did not promote GR–GRE interaction. These results demonstrate that induction and maintenance of nucleosome positioning is an active process that requires constant ‘pressure’ of agonist–GR‐recruited chromatin‐modifying factor(s) rather than GR–DNA binding itself.

[1]  P. Chambon,et al.  Folding of the DNA double helix in chromatin-like structures from simian virus 40. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[2]  E. Bradbury,et al.  Histone acetylation reduces nucleosome core particle linking number change , 1989, Cell.

[3]  M. C. Huber,et al.  Genomic position effects lead to an inefficient reorganization of nucleosomes in the 5'-regulatory region of the chicken lysozyme locus in transgenic mice. , 1996, Nucleic acids research.

[4]  C. Allis,et al.  The language of covalent histone modifications , 2000, Nature.

[5]  C. Woodcock,et al.  Nucleosome positioning by the winged helix transcription factor HNF3. , 1998, Genes & development.

[6]  M. Dunaway,et al.  Modeling transcriptional regulation using microinjection into Xenopus oocytes. , 1999, Methods.

[7]  T. Richmond,et al.  Positioning and stability of nucleosomes on MMTV 3'LTR sequences. , 1998, Journal of molecular biology.

[8]  M. Beato,et al.  Antiprogestins prevent progesterone receptor binding to hormone responsive elements in vivo. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[9]  W. Pennie,et al.  The Position and Length of the Steroid-Dependent Hypersensitive Region in the Mouse Mammary Tumor Virus Long Terminal Repeat Are Invariant despite Multiple Nucleosome B Frames , 1998, Molecular and Cellular Biology.

[10]  T. Archer,et al.  Chromatin remodelling by the glucocorticoid receptor requires the BRG1 complex , 1998, Nature.

[11]  K. Zaret,et al.  An active tissue-specific enhancer and bound transcription factors existing in a precisely positioned nucleosomal array , 1993, Cell.

[12]  A. Wolffe,et al.  Characterization of a chromatin remodelling activity in Xenopus oocytes. , 1999, European journal of biochemistry.

[13]  R. Kingston,et al.  ATP-dependent remodeling and acetylation as regulators of chromatin fluidity. , 1999, Genes & development.

[14]  Myles Brown,et al.  Cofactor Dynamics and Sufficiency in Estrogen Receptor–Regulated Transcription , 2000, Cell.

[15]  A. Wolffe,et al.  Targeting of N‐CoR and histone deacetylase 3 by the oncoprotein v‐ErbA yields a chromatin infrastructure‐dependent transcriptional repression pathway , 2000, The EMBO journal.

[16]  A. Wolffe,et al.  DNA methylation and histone deacetylation in the control of gene expression: basic biochemistry to human development and disease. , 2000, Gene expression.

[17]  G. Rousseau,et al.  Structure-activity relationships for glucocorticoids-I. Determination of receptor binding and biological activity. , 1977, Journal of steroid biochemistry.

[18]  E. Baulieu,et al.  RU486 (mifepristone): mechanisms of action and clinical uses. , 1997, Annual review of medicine.

[19]  G. Hager.  Understanding nuclear receptor function: from DNA to chromatin to the interphase nucleus. , 2001, Progress in nucleic acid research and molecular biology.

[20]  T. Perlmann,et al.  Specific glucocorticoid receptor binding to DNA reconstituted in a nucleosome. , 1988, The EMBO journal.

[21]  M. Beato,et al.  Transcription factor access to chromatin. , 1997, Nucleic acids research.

[22]  P. Chambon,et al.  Type II antagonists impair the DNA binding of steroid hormone receptors without affecting dimerization , 1993, The Journal of Steroid Biochemistry and Molecular Biology.

[23]  P. Chambon,et al.  The hormone-binding domains of the estrogen and glucocorticoid receptors contain an inducible transcription activation function , 1988, Cell.

[24]  T. Edlund,et al.  Sequence-specific interactions of nuclear factors with the insulin gene enhancer , 1986, Cell.

[25]  T. Reich,et al.  Nucleocytoplasmic Trafficking of Steroid-free Glucocorticoid Receptor* , 1999, The Journal of Biological Chemistry.

[26]  G. Hager,et al.  Transcription factor access is mediated by accurately positioned nucleosomes on the mouse mammary tumor virus promoter , 1991, Molecular and cellular biology.

[27]  T. Richmond,et al.  Crystal structure of the nucleosome core particle at 2.8 Å resolution , 1997, Nature.

[28]  M. Beato,et al.  Assembly of MMTV promoter minichromosomes with positioned nucleosomes precludes NF1 access but not restriction enzyme cleavage. , 1998, Nucleic acids research.

[29]  P. Eriksson,et al.  Protein-protein contacts in the glucocorticoid receptor homodimer influence its DNA binding properties. , 1990, The Journal of biological chemistry.

[30]  A. Wolffe,et al.  Superhelical stress and nucleosome‐mediated repression of 5S RNA gene transcription in vitro. , 1991, The EMBO journal.

[31]  J. Mymryk,et al.  Dissection of progesterone receptor-mediated chromatin remodeling and transcriptional activation in vivo. , 1995, Genes & development.

[32]  Keith R. Yamamoto,et al.  Reversible and persistent changes in chromatin structure accompany activation of a glucocorticoid-dependent enhancer element , 1984, Cell.

[33]  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.

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

[35]  R. Simpson,et al.  Nucleosome positioning: occurrence, mechanisms, and functional consequences. , 1991, Progress in nucleic acid research and molecular biology.

[36]  H. Richard-Foy,et al.  Sequence‐specific positioning of nucleosomes over the steroid‐inducible MMTV promoter. , 1987, The EMBO journal.

[37]  J. Workman,et al.  Promoter targeting and chromatin remodeling by the SWI/SNF complex. , 2000, Current opinion in genetics & development.

[38]  K. Yamamoto,et al.  Two signals mediate hormone‐dependent nuclear localization of the glucocorticoid receptor. , 1987, The EMBO journal.

[39]  R. Tjian,et al.  Orchestrated response: a symphony of transcription factors for gene control. , 2000, Genes & development.

[40]  M. Beato,et al.  Nucleosome positioning modulates accessibility of regulatory proteins to the mouse mammary tumor virus promoter , 1990, Cell.

[41]  G. Almouzni,et al.  Hormone activation induces nucleosome positioning in vivo , 2000, The EMBO journal.

[42]  K. Zaret,et al.  An early developmental transcription factor complex that is more stable on nucleosome core particles than on free DNA. , 1999, Molecular cell.

[43]  J. McNally,et al.  The glucocorticoid receptor: rapid exchange with regulatory sites in living cells. , 2000, Science.

[44]  M. Perricaudet,et al.  Receptors bound to antiprogestin form abortive complexes with hormone responsive elements , 1988, Nature.

[45]  G. Ryffel,et al.  Two types of antiprogestins identified by their differential action in transcriptionally active extracts from T47D cells. , 1991, Nucleic acids research.

[46]  G. Hager,et al.  Nucleosomes reconstituted in vitro on mouse mammary tumor virus B region DNA occupy multiple translational and rotational frames. , 1995, Biochemistry.

[47]  A. Wolffe,et al.  Review: chromatin structural features and targets that regulate transcription. , 2000, Journal of structural biology.

[48]  S. Elgin,et al.  Nucleosome positioning and gene regulation , 1994, Journal of cellular biochemistry.

[49]  M. Stallcup,et al.  GRIP1, a novel mouse protein that serves as a transcriptional coactivator in yeast for the hormone binding domains of steroid receptors. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[50]  G. Hager,et al.  Nucleosome positioning on the MMTV LTR results from the frequency-biased occupancy of multiple frames. , 1995, Genes & development.

[51]  A. Wolffe,et al.  Replication-coupled chromatin assembly is required for the repression of basal transcription in vivo. , 1993, Genes & development.

[52]  Dimitris Thanos,et al.  Ordered Recruitment of Chromatin Modifying and General Transcription Factors to the IFN-β Promoter , 2000, Cell.

[53]  R. Kingston,et al.  BRG-1 Is Recruited to Estrogen-Responsive Promoters and Cooperates with Factors Involved in Histone Acetylation , 2000, Molecular and Cellular Biology.

[54]  W. Gilbert,et al.  A new method for sequencing DNA. , 1977, Proceedings of the National Academy of Sciences of the United States of America.