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Originally published as MBC in Press, 10.1091/mbc.E03-06-0444 on January 12, 2004

Vol. 15, Issue 4, 1568-1579, April 2004

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TopBP1 and ATR Colocalization at Meiotic Chromosomes: Role of TopBP1/Cut5 in the Meiotic Recombination Checkpoint

David Perera * {dagger}, Livia Perez-Hidalgo {ddagger} {dagger}, Peter B. Moens §, Kaarina Reini ¶, Nicholas Lakin ||, Juhani E. Syväoja ¶ #, Pedro A. San-Segundo {ddagger} **, and Raimundo Freire * **

* Unidad de Investigación, Hospital Universitario de Canarias, Ofra s/n, La Cuesta, 38320 Tenerife, Spain; {ddagger} Centro de Investigación del Cáncer, CSIC/University of Salamanca, Campus Unamuno, 37007 Salamanca, Spain; § Department of Biology, York University, Downsview, Ontario, M3J 1P3 Canada; Biocenter Oulu and Department of Biochemistry, University of Oulu, FIN-90014 Finland; || Division of Molecular Genetics, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, United Kingdom; and # Department of Biology, University of Joensuu, FIN-80101 Joensuu, Finland

Submitted June 27, 2003; Revised December 15, 2003; Accepted December 15, 2003
Monitoring Editor: Mark Solomon

Mammalian TopBP1 is a BRCT domain–containing protein whose function in mitotic cells is linked to replication and DNA damage checkpoint. Here, we study its possible role during meiosis in mice. TopBP1 foci are abundant during early prophase I and localize mainly to histone {gamma}-H2AX–positive domains, where DNA double–strand breaks (required to initiate recombination) occur. Strikingly, TopBP1 showed a pattern almost identical to that of ATR, a PI3K-like kinase involved in mitotic DNA damage checkpoint. In the synapsis-defective Fkbp6-/- mouse, TopBP1 heavily stains unsynapsed regions of chromosomes. We also tested whether Schizosaccharomyces pombe Cut5 (the TopBP1 homologue) plays a role in the meiotic recombination checkpoint, like spRad3, the ATR homologue. Indeed, we found that a cut5 mutation suppresses the checkpoint-dependent meiotic delay of a meiotic recombination defective mutant, indicating a direct role of the Cut5 protein in the meiotic checkpoint. Our findings suggest that ATR and TopBP1 monitor meiotic recombination and are required for activation of the meiotic recombination checkpoint.

Abbreviations used: SC, synaptonemal complex.; DSBs, DNA double-strand breaks.; PIKK, phosphatidyl-inositol 3-kinase-like protein kinase.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03–06–0444. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03-06-0444.

{dagger} These authors contributed equally to this work.

** Corresponding authors. E-mail addresses: pedross{at}usal.es and rfreire{at}hecit.es.




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