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Genetic analysis ofDNA repair in thehyperthermophilicarchaeon,Thermococcuskodakaraensis Ryosuke FujikaneSonoko IshinoYoshizumi Ishino2010 год

Genetic analysis of DNA repair in the hyperthermophilic archaeon, Thermococcus kodakaraensis
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Аннотация: Extensive biochemical and structural analyses have been performed on the putative DNA repair proteins of hyperthermophilic archaea, in contrast to the few genetic analyses of the genes encoding these proteins. Accordingly, little is known about the repair pathways used by archaeal cells at high temperature. Here, we attempted to disrupt the genes encoding the potential repair proteins in the genome of the hyperthermophilic archaeon Thermococcus kodakaraensis. We succeeded in isolating null mutants of the hjc, hef, hjm, xpb, and xpd genes, but not the radA, rad50, mre11, herA, nurA, and xpg/fen1 genes. Phenotypic analyses of the gene-disrupted strains showed that the xpb and xpd null mutants are only slightly sensitive to ultraviolet (UV) irradiation, methyl methanesulfonate (MMS) and mitomycin C (MMC), as compared with the wild-type strain. The hjm null mutant showed sensitivity specifically to mitomycin C. On the other hand, the null mutants of the hjc gene lacked increasing sensitivity to any type of DNA damage. The Hef protein is particularly important for maintaining genome homeostasis, by functioning in the repair of a wide variety of DNA damage in T. kodakaraensis cells. Deletion of the entire hef gene or of the segments encoding either its nuclease or helicase domain produced similar phenotypes. The high sensitivity of the Δhef mutants to MMC suggests that Hef performs a critical function in the repair process of DNA interstrand cross-links. These damage-sensitivity profiles suggest that the archaeal DNA repair system has processes depending on repair-related proteins different from those of eukaryotic and bacterial DNA repair systems using homologous repair proteins analyzed here.
Год издания: 2010
Авторы: Ryosuke Fujikane, Sonoko Ishino, Yoshizumi Ishino, Patrick Forterre
Издательство: Genetics Society of Japan
Источник: Genes & Genetic Systems
Ключевые слова: DNA Repair Mechanisms, Bacterial Genetics and Biotechnology, Enzyme Structure and Function
Другие ссылки: Genes & Genetic Systems (PDF)
Genes & Genetic Systems (HTML)
PubMed (HTML)