The Cdc45/RecJ-like protein forms a complex with GINS and MCM, and is important for DNA replication in Thermococcus kodakarensis

Mariko Nagata, Sonoko Ishino, Takeshi Yamagami, Hiromi Ogino, Jan Robert Simons, Tamotsu Kanai, Haruyuki Atomi, Yoshizumi Ishino

研究成果: Contribution to journalArticle査読

14 被引用数 (Scopus)

抄録

The archaeal minichromosome maintenance (MCM) has DNA helicase activity, which is stimulated by GINS in several archaea. In the eukaryotic replicative helicase complex, Cdc45 forms a complex with MCM and GINS, named as CMG (Cdc45-MCM-GINS). Cdc45 shares sequence similarity with bacterial RecJ. A Cdc45/RecJ-like protein from Thermococcus kodakarensis shows a bacterial RecJ-like exonuclease activity, which is stimulated by GINS in vitro. Therefore, this archaeal Cdc45/RecJ is designated as GAN, from GINS-associated nuclease. In this study, we identified the CMG-like complex in T. kodakarensis cells. The GANGINS complex stimulated the MCM helicase, but MCM did not affect the nuclease activity of GAN in vitro. The gene disruption analysis showed that GAN was non-essential for its viability but the Agan mutant did not grow at 93 C. Furthermore, the Agan mutant showed a clear retardation in growth as compared with the parent cells under optimal conditions at 85 C. These deficiencies were recovered by introducing the gan gene encoding the nuclease deficient GAN protein back to the genome. These results suggest that the replicative helicase complex without GAN may become unstable and ineffective in replication fork progression. The nuclease activity of GAN is not related to the growth defects of the δgan mutant cells.

本文言語英語
ページ(範囲)10693-10705
ページ数13
ジャーナルNucleic acids research
45
18
DOI
出版ステータス出版済み - 10 1 2017

All Science Journal Classification (ASJC) codes

  • Genetics

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