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Molecular and Cellular Biology, July 2006, p. 4843-4852, Vol. 26, No. 13
0270-7306/06/$08.00+0     doi:10.1128/MCB.02267-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The N-Terminal Noncatalytic Region of Xenopus RecQ4 Is Required for Chromatin Binding of DNA Polymerase {alpha} in the Initiation of DNA Replication

Kumiko Matsuno, Maya Kumano, Yumiko Kubota, Yoshitami Hashimoto, and Haruhiko Takisawa*

Department of Biological Sciences, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan

Received 26 November 2005/ Returned for modification 27 December 2005/ Accepted 14 April 2006

Recruitment of DNA polymerases onto replication origins is a crucial step in the assembly of eukaryotic replication machinery. A previous study in budding yeast suggests that Dpb11 controls the recruitment of DNA polymerases {alpha} and {varepsilon} onto the origins. Sld2 is an essential replication protein that interacts with Dpb11, but no metazoan homolog has yet been identified. We isolated Xenopus RecQ4 as a candidate Sld2 homolog. RecQ4 is a member of the metazoan RecQ helicase family, and its N-terminal region shows sequence similarity with Sld2. In Xenopus egg extracts, RecQ4 is essential for the initiation of DNA replication, in particular for chromatin binding of DNA polymerase {alpha}. An N-terminal fragment of RecQ4 devoid of the helicase domain could rescue the replication activity of RecQ4-depleted extracts, and antibody against the fragment inhibited DNA replication and chromatin binding of the polymerase. Further, N-terminal fragments of RecQ4 physically interacted with Cut5, a Xenopus homolog of Dpb11, and their ability to bind to Cut5 closely correlated with their ability to rescue the replication activity of the depleted extracts. Our data suggest that RecQ4 performs an essential role in the assembly of replication machinery through interaction with Cut5 in vertebrates.

* Corresponding author. Mailing address: Department of Biological Sciences, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan. Phone: 81-6-6850-6762. Fax: 81-6-6850-6762. E-mail: takisawa{at}bio.sci.osaka-u.ac.jp.

Molecular and Cellular Biology, July 2006, p. 4843-4852, Vol. 26, No. 13
0270-7306/06/$08.00+0     doi:10.1128/MCB.02267-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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