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J. Biol. Chem., Vol. 278, Issue 3, 1924-1931, January 17, 2003
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From the The Ku70-Ku80 heterodimer is a conserved protein
complex essential for the non-homologous end-joining pathway. Ku
proteins are also involved in telomere maintenance, although their
precise roles remain to be elucidated. In fission yeast,
pku70+, the gene encoding the Ku70 homologue,
has been reported. Here we report the identification and
characterization of pku80+, the gene encoding
Ku80. Both pku70+ and
pku80+ are essential for efficient
non-homologous end-joining. We also found that the pku70
and pku80 mutants are sensitive to methyl methanesulfonate
and hydroxyurea, suggesting their roles in the S phase. The
pku80 mutant shows telomere shortening and tandem amplification of a subtelomeric sequence but no defects in the telomere
position effect, as was previously reported for the pku70 mutant. By using the chromatin immunoprecipitation assay, we
demonstrated that Pku70 and Pku80 physically interact with telomeric
repeats and subtelomeric sequences. Interestingly, this telomere
association of Pku proteins is independent of Taz1, a telomeric
DNA-binding protein. We also showed that the Pku proteins do not
associate with ectopically integrated telomeric repeats in the internal region of circular chromosomes. These results indicate that the physical end of DNA is necessary for the localization of Pku80 at telomeres.
Telomeric DNA Ends Are Essential for the Localization of Ku
at Telomeres in Fission Yeast*
,,§¶
Graduate School of Bioscience and
Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta,
Midori-ku, Yokohama 226-8501 and § Graduate School of
Biostudies, Kyoto University, Kitashirakawa-Oiwakecho, Sakyo-ku,
Kyoto 606-8502, Japan
*
This work was supported by a Center of Excellence
grant (to F. I.), a grant-in-aid for Cancer Research from the Ministry
of Education, Culture, Sports, Science and Technology, Japan (to F. I.), a Health and Labor Sciences Research grant from the Ministry of Health, Labor and Welfare, Japan (to F. I.), and a grant-in-aid from the Ministry of Education, Culture, Sports, Science and
Technology, Japan (to J. K.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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