title: Identification of a mismatch-specific endonuclease in hyperthermophilic Archaea.
abstract: The common mismatch repair system processed by MutS and MutL and their homologs was identified in Bacteria and Eukarya. However, no evidence of a functional MutS/L homolog has been reported for archaeal organisms, and it is not known whether the mismatch repair system is conserved in Archaea. Here, we describe an endonuclease that cleaves double-stranded DNA containing a mismatched base pair, from the hyperthermophilic archaeon Pyrococcus furiosus The corresponding gene revealed that the activity originates from PF0012, and we named this enzyme Endonuclease MS (EndoMS) as the mismatch-specific Endonuclease. The sequence similarity suggested that EndoMS is the ortholog of NucS isolated from Pyrococcus abyssi, published previously. Biochemical characterizations of the EndoMS homolog from Thermococcus kodakarensis clearly showed that EndoMS specifically cleaves both strands of double-stranded DNA into 5'-protruding forms, with the mismatched base pair in the central position. EndoMS cleaves G/T, G/G, T/T, T/C and A/G mismatches, with a more preference for G/T, G/G and T/T, but has very little or no effect on C/C, A/C and A/A mismatches. The discovery of this endonuclease suggests the existence of a novel mismatch repair process, initiated by the double-strand break generated by the EndoMS endonuclease, in Archaea and some Bacteria.
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, Fukuoka 812-8581, Japan sonoko@agr.kyushu-u.ac.jp.
authors.forename: Sonoko
authors.initials: S
authors.lastname: Ishino
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, Fukuoka 812-8581, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Nishi
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, Fukuoka 812-8581, Japan.
authors.forename: Soichiro
authors.initials: S
authors.lastname: Oda
authors.affiliation: Takara Bio Inc., Nojihigashi 7-4-38, Kusatsu, Shiga 525-0058, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Uemori
authors.affiliation: Takara Bio Inc., Nojihigashi 7-4-38, Kusatsu, Shiga 525-0058, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Sagara
authors.affiliation: Takara Bio Inc., Nojihigashi 7-4-38, Kusatsu, Shiga 525-0058, Japan.
authors.forename: Nariaki
authors.initials: N
authors.lastname: Takatsu
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, Fukuoka 812-8581, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yamagami
authors.affiliation: Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Shirai
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, Fukuoka 812-8581, Japan ishino@agr.kyushu-u.ac.jp.
authors.forename: Yoshizumi
authors.initials: Y
authors.lastname: Ishino
mesh_terms: D019843:Archaeal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D020137:Base Pair Mismatch; D053837:DNA Cleavage; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D020027:Pyrococcus furiosus / Q000201:enzymology; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D019843:Archaeal Proteins; D018809:Proliferating Cell Nuclear Antigen; D011994:Recombinant Proteins; D004706:Endodeoxyribonucleases