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Molecular and Cellular Biology, April 2004, p. 3036-3047, Vol. 24, No. 7
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.7.3036-3047.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Chromatin-Mediated Restriction of Nuclear Factor 1/CTF Binding in a Repressed and Hormone-Activated Promoter In Vivo

Sergey Belikov, Carolina Åstrand, Per-Henrik Holmqvist, and Örjan Wrange*

Department of Cell and Molecular Biology, The Medical Nobel Institute, Karolinska Institutet, SE-17177 Stockholm, Sweden

Received 16 December 2003/ Accepted 7 January 2004

Mouse mammary tumor virus (MMTV) promoter-driven transcription is induced by glucocorticoid hormone via binding of the glucocorticoid receptor (GR). The MMTV promoter also harbors a binding site for nuclear factor 1 (NF1). NF1 and GR were expressed in Xenopus oocytes; this revealed GR-NF1 cooperativity both in terms of DNA binding and chromatin remodeling but not transcription. A fraction of NF1 sites were occupied in a hormone-dependent fashion, but a significant and NF1 concentration-dependent fraction were constitutively bound. Activation of the MMTV promoter resulted in an ~50-fold increase in the NF1 accessibility for its DNA site. The hormone-dependent component of NF1 binding was dissociated by addition of a GR antagonist; however, the antagonist RU486, which supports partial GR-DNA binding, also maintained partial NF1 binding. Hence GR-NF1 cooperativity is independent of agonist-driven chromatin remodeling. NF1 induced the formation of a micrococcal-nuclease-resistant protein-DNA complex containing the DNA segment from -185 to -55, the MMTV enhanceosome. Coexpression of NF1 and Oct1 resulted in a significant stimulation of hormone-induced MMTV transcription and also in increased basal transcription. We propose that hormone-independent NF1 binding may be involved in maintaining transcriptional competence and establishment of tissue-specific gene networks.

* Corresponding author. Mailing address: Dept. of Cell and Molecular Biology, The Medical Nobel Institute, Box 285, Karolinska Institutet, SE-17177 Stockholm, Sweden. Phone: 46 8 728 7373. Fax: 46 8 31 35 29. E-mail: orjan.wrange{at}cmb.ki.se.

Molecular and Cellular Biology, April 2004, p. 3036-3047, Vol. 24, No. 7
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.7.3036-3047.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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