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Molecular and Cellular Biology, September 2003, p. 6553-6563, Vol. 23, No. 18
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.18.6553-6563.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of the Schizosaccharomyces pombe nbs1+ Gene Involved in DNA Repair and Telomere Maintenance

Masaru Ueno,1* Tomofumi Nakazaki,2 Yufuko Akamatsu,2,3 Kikuo Watanabe,1 Kazunori Tomita,1 Howard D. Lindsay,4 Hideo Shinagawa,3 and Hiroshi Iwasaki2

Department of Chemistry, Faculty of Science, Shizuoka University, Shizuoka 422-8529,1 Graduate School of Integrated Science, Yokohama City University, Tsurumi-ku, Yokohama, Kanagawa 230-0045,2 Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan,3 Genome Damage and Stability Centre, University of Sussex, Brighton BN1 9RQ, United Kingdom4

Received 10 April 2003/ Returned for modification 17 May 2003/ Accepted 6 June 2003

The human MRN complex is a multisubunit nuclease that is composed of Mre11, Rad50, and Nbs1 and is involved in homologous recombination and DNA damage checkpoints. Mutations of the MRN genes cause genetic disorders such as Nijmegen breakage syndrome. Here we identified a Schizosaccharomyces pombe nbs1+ homologue by screening for mutants with mutations that caused methyl methanesulfonate (MMS) sensitivity and were synthetically lethal with the rad2{Delta} mutation. Nbs1 physically interacts with the C-terminal half of Rad32, the Schizosaccharomyces pombe Mre11 homologue, in a yeast two-hybrid assay. nbs1 mutants showed sensitivities to {gamma}-rays, UV, MMS, and hydroxyurea and displayed telomere shortening similar to the characteristics of rad32 and rad50 mutants. nbs1, rad32, and rad50 mutant cells were elongated and exhibited abnormal nuclear morphology. These findings indicate that S. pombe Nbs1 forms a complex with Rad32-Rad50 and is required for homologous recombination repair, telomere length regulation, and the maintenance of chromatin structure. Amino acid sequence features and some characteristics of the DNA repair function suggest that the S. pombe Rad32-Rad50-Nbs1 complex has functional similarity to the corresponding MRN complexes of higher eukaryotes. Therefore, S. pombe Nbs1 will provide an additional model system for studying the molecular function of the MRN complex associated with genetic diseases.

* Corresponding author. Mailing address: Department of Chemistry, Faculty of Science, Shizuoka University, 836 Oya, Shizuoka 422-8529, Japan. Phone: 81 54 238 4762. Fax: 81 54 237 3384. E-mail: scmueno{at}ipc.shizuoka.ac.jp.

Molecular and Cellular Biology, September 2003, p. 6553-6563, Vol. 23, No. 18
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.18.6553-6563.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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