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Molecular and Cellular Biology, January 2005, p. 797-807, Vol. 25, No. 2
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.2.797-807.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Histone Chaperone TAF-I/SET/INHAT Is Required for Transcription In Vitro of Chromatin Templates

Matthew J. Gamble,1,2 Hediye Erdjument-Bromage,1 Paul Tempst,1 Leonard P. Freedman,3,{dagger} and Robert P. Fisher1*

Molecular Biology Program,1 Cell Biology Program, Memorial Sloan-Kettering Cancer Center,3 Programs in Biochemistry, Cell and Molecular Biology, Cornell University Graduate School of Medical Sciences, New York, New York2

Received 23 July 2004/ Returned for modification 2 September 2004/ Accepted 18 October 2004

To uncover factors required for transcription by RNA polymerase II on chromatin, we fractionated a mammalian cell nuclear extract. We identified the histone chaperone TAF-I (also known as INHAT [inhibitor of histone acetyltransferase]), which was previously proposed to repress transcription, as a potent activator of chromatin transcription responsive to the vitamin D3 receptor or to Gal4-VP16. TAF-I associates with chromatin in vitro and can substitute for the related protein NAP-1 in assembling chromatin onto cloned DNA templates in cooperation with the remodeling enzyme ATP-dependent chromatin assembly factor (ACF). The chromatin assembly and transcriptional activation functions are distinct, however, and can be dissociated temporally. Efficient transcription of chromatin assembled with TAF-I still requires the presence of TAF-I during the polymerization reaction. Conversely, TAF-I cannot stimulate transcript elongation when added after the other factors necessary for assembly of a preinitiation complex on naked DNA. Thus, TAF-I is required to facilitate transcription at a step after chromatin assembly but before transcript elongation.

* Corresponding author. Mailing address: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Ave., New York, NY 10021. Phone: (212) 639-8912. Fax: (212) 717-3317. E-mail: r-fisher{at}ski.mskcc.org.

{dagger} Present address: Department of Molecular Endocrinology, Merck Research Laboratories, West Point, PA 19486.

Molecular and Cellular Biology, January 2005, p. 797-807, Vol. 25, No. 2
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.2.797-807.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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