Binding of Acf1 to DNA involves a WAC motif and is important for ACF-mediated chromatin assembly.
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.
Section of Molecular Biology, University of California, San Diego, La Jolla 92093-0347, USA.
Dmitry V
DV
Fyodorov
James T
JT
Kadonaga
D000251:Adenosine Triphosphatases / Q000378:metabolism; D020816:Amino Acid Motifs; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004278:DNA, Superhelical; D004330:Drosophila; D029721:Drosophila Proteins; D007313:Insecta; D008957:Models, Genetic; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism*
C120419:Acf protein, Drosophila; D002843:Chromatin; D018076:DNA, Complementary; D004278:DNA, Superhelical; D029721:Drosophila Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; D013395:Sucrose; D004247:DNA; D000251:Adenosine Triphosphatases