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Papers In Press, published online ahead of print March 14, 2006
Department of Biochemistry, Miyazaki Medical College, University of Miyazaki, Kiyotake, Miyazaki 889-1692
Corresponding Author: tnakayam{at}med.miyazaki-u.ac.jp
Anti silencing function 1 (Asf1), a well conserved protein from yeast to human, acts as a histone chaperone and is predicted to participate in a variety of chromatin mediated cellular processes. To investigate the physiological role of vertebrate Asf1 in vivo, we generated a conditional Asf1 deficient mutant from chicken DT40 cells. Induction of Asf1-depletion resulted in the accumulation of cells in S phase with decreased DNA replication and increased mitotic aberrancy forming multipolar spindles, leading to cell death. In addition, nascent chromatin in Asf1 depleted cells showed increased nuclease sensitivity, indicating impaired nucleosome assembly during DNA replication. Complementation analyses revealed that the functional domain of Asf1 for cell viability was confined to the N-terminal core domain (1-155 amino acids) that is a binding platform for histones H3/H4, CAF-1p60 and HIRA, while Asf1 mutant proteins, abolishing binding abilities with both p60 and HIRA, exhibit no effect on viability. These results, together, indicate that the vertebrate Asf1 plays a crucial role in replication coupled chromatin assembly, cell cycle progression and cellular viability, and provides a clue of possible role in a CAF-1 and HIRA independent chromatin modulating process for cell proliferation.
J. Biol. Chem, 10.1074/jbc.M511590200
Submitted on October 26, 2005
Revised on March 14, 2006
Accepted on March 14, 2006
Asf1 is required for viability and chromatin assembly during DNA replication in vertebrate cells
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