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Molecular and Cellular Biology, November 2004, p. 9557-9567, Vol. 24, No. 21
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.21.9557-9567.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Fission Yeast Dna2 Is Required for Generation of the Telomeric Single-Strand Overhang

Kazunori Tomita,1,{dagger} Tatsuya Kibe,1 Ho-Young Kang,2 Yeon-Soo Seo,2 Masahiro Uritani,1 Takashi Ushimaru,3 and Masaru Ueno1*

Department of Chemistry,1 Department of Biology, Shizuoka University, Shizuoka, Japan,3 Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yusung-Ku, Daejeon, South Korea2

Received 22 April 2004/ Returned for modification 18 May 2004/ Accepted 11 August 2004

It has been suggested that the Schizosaccharomyces pombe Rad50 (Rad50-Rad32-Nbs1) complex is required for the resection of the C-rich strand at telomere ends in taz1-d cells. However, the nuclease-deficient Rad32-D25A mutant can still resect the C-rich strand, suggesting the existence of a nuclease that resects the C-rich strand. Here, we demonstrate that a taz1-d dna2-2C double mutant lost the G-rich overhang at a semipermissive temperature. The amount of G-rich overhang in S phase in the dna2-C2 mutant was lower than that in wild-type cells at the semipermissive temperature. Dna2 bound to telomere DNA in a chromatin immunoprecipitation assay. Moreover, telomere length decreased with each generation after shift of the dna2-2C mutant to the semipermissive temperature. These results suggest that Dna2 is involved in the generation of G-rich overhangs in both wild-type cells and taz1-d cells. The dna2-C2 mutant was not gamma ray sensitive at the semipermissive temperature, suggesting that the ability to process double-strand break (DSB) ends was not affected in the dna2-C2 mutant. Our results reveal that DSB ends and telomere ends are processed by different mechanisms.

* 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.

{dagger} Present address: Telomere Biology Laboratory, Cancer Research UK, London EC2A 3PX, United Kingdom.

Molecular and Cellular Biology, November 2004, p. 9557-9567, Vol. 24, No. 21
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.21.9557-9567.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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