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Molecular and Cellular Biology, September 2002, p. 6344-6353, Vol. 22, No. 18
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.18.6344-6353.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Binding of Acf1 to DNA Involves a WAC Motif and Is Important for ACF-Mediated Chromatin Assembly

Dmitry V. Fyodorov and James T. Kadonaga*

Section of Molecular Biology, University of California, San Diego, La Jolla, California 92093-0347

Received 1 May 2002/ Accepted 11 June 2002

ACF is a chromatin-remodeling complex that catalyzes the ATP-dependent assembly of periodic nucleosome arrays. This reaction utilizes the energy of ATP hydrolysis by ISWI, the smaller of the two subunits of ACF. Acf1, the large subunit of ACF, is essential for the full activity of the complex. We performed a systematic mutational analysis of Acf1 to elucidate the functions of specific subregions of the protein. These studies revealed DNA- and ISWI-binding regions that are important for the chromatin assembly and ATPase activities of ACF. The DNA-binding region of Acf1 includes a WAC motif, which is necessary for the efficient binding of ACF complex to DNA. The interaction of Acf1 with ISWI requires a DDT domain, which has been found in a variety of transcription and chromatin-remodeling factors. Chromatin assembly by ACF is also impaired upon mutation of an acidic region in Acf1, which may interact with histones during the deposition process. Lastly, we observed modest chromatin assembly defects on mutation of other conserved sequence motifs. Thus, Acf1 facilitates chromatin assembly via an N-terminal DNA-binding region with a WAC motif, a central ISWI-binding segment with a DDT domain, and a C-terminal region with an acidic stretch, a WAKZ motif, PHD fingers, and bromodomain.

* Corresponding author. Mailing address: Section of Molecular Biology, 0347, Pacific Hall, Room 2212B, University of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093-0347. Phone: (858) 534-4608. Fax: (858) 534-0555. E-mail: jkadonaga{at}ucsd.edu.

Molecular and Cellular Biology, September 2002, p. 6344-6353, Vol. 22, No. 18
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.18.6344-6353.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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