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Molecular and Cellular Biology, September 2008, p. 5764-5776, Vol. 28, No. 18
0270-7306/08/$08.00+0     doi:10.1128/MCB.01262-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Different Functional Modes of p300 in Activation of RNA Polymerase III Transcription from Chromatin Templates{triangledown}

Claudia Mertens{dagger} and Robert G. Roeder*

Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, New York 10021

Received 13 July 2007/ Returned for modification 31 August 2007/ Accepted 7 July 2008

Transcriptional coactivators that regulate the activity of human RNA polymerase III (Pol III) in the context of chromatin have not been reported. Here, we describe a completely defined in vitro system for transcription of a human tRNA gene assembled into a chromatin template. Transcriptional activation and histone acetylation in this system depend on recruitment of p300 by general initiation factor TFIIIC, thus providing a new paradigm for recruitment of histone-modifying coactivators. Beyond its role as a chromatin-modifying factor, p300 displays an acetyltransferase-independent function at the level of preinitiation complex assembly. Thus, direct interaction of p300 with TFIIIC stabilizes binding of TFIIIC to core promoter elements and results in enhanced transcriptional activity on histone-free templates. Additional studies show that p300 is recruited to the promoters of actively transcribed tRNA and U6 snRNA genes in vivo. These studies identify TFIIIC as a recruitment factor for p300 and thus may have important implications for the emerging concept that tRNA genes or TFIIIC binding sites act as chromatin barriers to prohibit spreading of silenced heterochromatin domains.

* Corresponding author. Mailing address: Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-7601. Fax: (212) 327-7949. E-mail: roeder{at}rockefeller.edu

{triangledown} Published ahead of print on 21 July 2008.

{dagger} Present address: Laboratory of Molecular Cell Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10021.

Molecular and Cellular Biology, September 2008, p. 5764-5776, Vol. 28, No. 18
0270-7306/08/$08.00+0     doi:10.1128/MCB.01262-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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