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Molecular and Cellular Biology, October 2001, p. 6574-6584, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6574-6584.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Interaction between the Drosophila CAF-1 and ASF1 Chromatin Assembly Factors

Jessica K. Tyler,1,2,* Kimberly A. Collins,1,dagger Jayashree Prasad-Sinha,3 Elizabeth Amiott,2 Michael Bulger,1,Dagger Peter J. Harte,3 Ryuji Kobayashi,4 and James T. Kadonaga1,*

Section of Molecular Biology, University of California, San Diego, La Jolla, California 92093-03471; Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, Denver, Colorado 802622; Department of Genetics, School of Medicine, Case Western Reserve University, Cleveland, Ohio 441063; and Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 117244

Received 4 April 2001/Returned for modification 17 May 2001/Accepted 6 July 2001

The assembly of newly synthesized DNA into chromatin is essential for normal growth, development, and differentiation. To gain a better understanding of the assembly of chromatin during DNA synthesis, we identified, cloned, and characterized the 180- and 105-kDa polypeptides of Drosophila chromatin assembly factor 1 (dCAF-1). The purified recombinant p180+p105+p55 dCAF-1 complex is active for DNA replication-coupled chromatin assembly. Furthermore, we have established that the putative 75-kDa polypeptide of dCAF-1 is a C-terminally truncated form of p105 that does not coexist in dCAF-1 complexes containing the p105 subunit. The analysis of native and recombinant dCAF-1 revealed an interaction between dCAF-1 and the Drosophila anti-silencing function 1 (dASF1) component of replication-coupling assembly factor (RCAF). The binding of dASF1 to dCAF-1 is mediated through the p105 subunit of dCAF-1. Consistent with the interaction between dCAF-1 p105 and dASF1 in vitro, we observed that dASF1 and dCAF-1 p105 colocalized in vivo in Drosophila polytene chromosomes. This interaction between dCAF-1 and dASF1 may be a key component of the functional synergy observed between RCAF and dCAF-1 during the assembly of newly synthesized DNA into chromatin.

* Corresponding author. Mailing address for Jessica K. Tyler: Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, 4200 East Ninth Ave., Denver, CO 80262. Mailing address for James T. Kadonaga: 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.

dagger Present address: Department of Biology, University of Washington, Seattle, WA 98195.

Dagger Present address: Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024.

Molecular and Cellular Biology, October 2001, p. 6574-6584, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6574-6584.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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