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M607432200v1
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Papers In Press, published online ahead of print December 22, 2006
J. Biol. Chem, 10.1074/jbc.M607432200
Submitted on August 4, 2006
Revised on December 21, 2006
Accepted on December 22, 2006

Tel2 is required for activation of the Mrc1-mediated replication checkpoint

Miho Shikata, Fuyuki Ishikawa, and Junko Kanoh

Graduate School of Biostudies, Kyoto University, Kyoto, Kyoto 606-8502

Corresponding Author: jkanoh{at}lif.kyoto-u.ac.jp

Proteins belonging to the Tel2/Rad-5/Clk-2 family are conserved among eukaryotes, and are involved in various cellular processes, such as cell proliferation, telomere maintenance, the biological clock, and the DNA damage checkpoint. However, the molecular mechanisms underlying the functions of these molecules remain largely unclear. Here, we report that in the fission yeast, Schizosaccharomyces pombe, Tel2 is required for efficient phosphorylation of Mrc1, a mediator of DNA replication checkpoint signaling, and for activation of Cds1, a replication checkpoint kinase, when DNA replication is blocked by hydroxyurea (HU). In fact, Tel2 is required for survival of replication fork arrest and for the replication checkpoint in cells lacking Chk1, another checkpoint kinase the role of which overlaps that of Cds1 in cell cycle arrest by replication block. In addition, Tel2 plays important roles in entry into S phase and in genome stability. Tel2 is essential for vegetative cell growth, and the tel2 strain accumulated cells with 1C DNA content after germination. In the absence of HU, Tel2 is vital in the mutant lacking Swi1, a component of the replication fork protection complex, and multiple Rad22 DNA repair foci were frequently observed in Tel2-repressed swi1 cells especially at S phase. In contrast, the cds1swi1 mutant did not show such lethality. These results indicate that S. pombe Tel2 plays important roles in the Mrc1-mediated replication checkpoint as well as in the Cds1-independent regulation of genome integrity.


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