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Molecular and Cellular Biology, April 2000, p. 2809-2817, Vol. 20, No. 8
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Dpb11 Controls the Association between DNA Polymerases alpha  and epsilon  and the Autonomously Replicating Sequence Region of Budding Yeast

Hiroshi Masumoto,1,2 Akio Sugino,2 and Hiroyuki Araki1,3,4,*

Division of Microbial Genetics, National Institute of Genetics,1 The Graduate University for Advanced Studies,3 and PRESTO, Japan Science and Technology Corporation,4 Shizuoka, and Research Institute for Microbial Diseases, Osaka University, Osaka,2 Japan

Received 15 November 1999/Returned for modification 13 January 2000/Accepted 28 January 2000

Dpb11 is required for chromosomal DNA replication and the S-phase checkpoint in Saccharomyces cerevisiae. Here, we report detection of a physical complex containing Dpb11 and DNA polymerase varepsilon  (Dpb11-Polvarepsilon complex). During the S phase of the cell cycle, Dpb11 associated preferentially with DNA fragments containing autonomously replicating sequences (ARSs), at the same time as Polvarepsilon associated with these fragments. Association of Dpb11 and Polvarepsilon with these fragments was mutually dependent, suggesting that the Dpb11-Polvarepsilon complex associates with the ARS. Moreover, Dpb11 was required for the association of Polalpha -primase with the fragments. Thus, it seems likely that association of the Dpb11-Polvarepsilon complex with the ARS fragments is required for the association of the Polalpha -primase complex. Hydroxyurea inhibits late-origin firing in S. cerevisiae, and the checkpoint genes, RAD53 and MEC1, are involved in this inhibition. In the presence of hydroxyurea at temperatures permissive for cell growth, Polvarepsilon in dpb11-1 cells associated with early- and late-origin fragments. In wild-type cells, however, it associated only with early-origin fragments. This indicates that Dpb11 may also be involved in the regulation of late-origin firing. Overall, these results suggest that Dpb11 controls the association between DNA polymerases alpha  and varepsilon  and the ARS.

* Corresponding author. Mailing address: Division of Microbial Genetics, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka 411-8540, Japan. Phone: (81) 559 81 6754. Fax: (81) 559 81 6762. E-mail: hiaraki{at}lab.nig.ac.jp.

Molecular and Cellular Biology, April 2000, p. 2809-2817, Vol. 20, No. 8
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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