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J. Biol. Chem., Vol. 283, Issue 3, 1601-1609, January 18, 2008

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The GINS Complex from Pyrococcus furiosus Stimulates the MCM Helicase Activity^*

Takehiro Yoshimochi, Ryosuke Fujikane¹, Miyuki Kawanami, Fujihiko Matsunaga, and Yoshizumi Ishino²

From the Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University and BIRD-Japan Science and Technology Agency, 6-10-1 Fukuoka 812-8581, Japan

Pyrococcus furiosus, a hyperthermophilic Archaea, has homologs of the eukaryotic MCM (mini-chromosome maintenance) helicase and GINS complex. The MCM and GINS proteins are both essential factors to initiate DNA replication in eukaryotic cells. Many biochemical characterizations of the replication-related proteins have been reported, but it has not been proved that the homologs of each protein are also essential for replication in archaeal cells. Here, we demonstrated that the P. furiosus GINS complex interacts with P. furiosus MCM. A chromatin immunoprecipitation assay revealed that the GINS complex is detected preferentially at the oriC region on Pyrococcus chromosomal DNA during the exponential growth phase but not in the stationary phase. Furthermore, the GINS complex stimulates both the ATPase and DNA helicase activities of MCM in vitro. These results strongly suggest that the archaeal GINS is involved in both the initiation and elongation processes of DNA replication in P. furiosus, as observed in eukaryotic cells.

Received for publication, September 12, 2007 , and in revised form, October 30, 2007.

^* This work was also supported in part by grants-in-aid from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (to Y. I. and F. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Present address: Institute de Genetique et Microbiologie, Universite Paris XI, ORSAY cedex, France.

² Supported by a research grant from the Human Frontier Science Program. To whom correspondence should be addressed: 6-10-1 Hakozaki, Higashiku, Fukuoka-shi, Fukuoka 812-8581, Japan. Fax: 81-92-642-3051; E-mail: ishino{at}agr.kyushu-u.ac.jp.

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