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Interactions between Mei4, Rec114, and other proteins required for meiotic DNA double-strand break formation in Saccharomyces cerevisiae

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Abstract

In most sexually reproducing organisms, meiotic recombination is initiated by DNA double-strand breaks (DSBs) formed by the Spo11 protein. In budding yeast, nine other proteins are also required for DSB formation, but the roles of these proteins and the interactions among them are poorly understood. We report further studies of the behaviors of these proteins. Consistent with other studies, we find that Mei4 and Rec114 bind to chromosomes from leptonema through early pachynema. Both proteins showed only limited colocalization with the meiotic cohesin subunit Rec8, suggesting that Mei4 and Rec114 associated preferentially with chromatin loops. Rec114 localization was independent of other DSB factors, but Mei4 localization was strongly dependent on Rec114 and Mer2. Systematic deletion analysis identified protein regions important for a previously described two-hybrid interaction between Mei4 and Rec114. We also report functional characterization of a previously misannotated 5′ coding exon of REC102. Sequences encoded in this exon are essential for DSB formation and for Rec102 interaction with Rec104, Spo11, Rec114, and Mei4. Finally, we also examined genetic requirements for a set of previously described two-hybrid interactions that can be detected only when the reporter strain is induced to enter meiosis. This analysis reveals new functional dependencies for interactions among the DSB proteins. Taken together, these studies support the view that Mei4, Rec114, and Mer2 make up a functional subgroup that is distinct from other subgroups of the DSB proteins: Spo11–Ski8, Rec102–Rec104, and Mre11–Rad50–Xrs2. These studies also suggest that an essential function of Rec102 and Rec104 is to connect Mei4 and Rec114 to Spo11.

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Acknowledgments

We are grateful to Dianna Fisk (Saccharomyces Genome Database) for alerting us to the presence of the 5′ exon in REC102. We thank Nancy Kleckner, Franz Klein, Bob Malone, and Shirleen Roeder for strains, plasmids and/or antibodies. This work was supported by NIH grant R01 GM58673 (to S.K.). M.J.N. is supported in part by a fellowship from the Human Frontiers Science Program. K.H. was supported in part by a Frank Lappin Horsfall, Jr. Fellowship and an Association for Women in Science Predoctoral Fellowship. S.K. is a Leukemia and Lymphoma Society Scholar.

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  1. Shohreh Maleki

    Present address: The Computational Medicine Group, Center for Molecular Medicine, Department of Medicine, Karolinska Institute, Stockholm, Sweden

  2. Kiersten A. Henderson

    Present address: Fred Hutchinson Cancer Research Center, Seattle, WA, USA

Authors and Affiliations

  1. Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Ave., New York, NY, 10021, USA

    Shohreh Maleki, Matthew J. Neale, Charanjit Arora, Kiersten A. Henderson & Scott Keeney

  2. Molecular Biology Program, Weill Graduate School of Medical Sciences, Cornell University, New York, NY, 10021, USA

    Charanjit Arora, Kiersten A. Henderson & Scott Keeney

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  1. Shohreh Maleki
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Correspondence to Scott Keeney.

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Communicated by E.A. Nigg

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Maleki, S., Neale, M.J., Arora, C. et al. Interactions between Mei4, Rec114, and other proteins required for meiotic DNA double-strand break formation in Saccharomyces cerevisiae . Chromosoma 116, 471–486 (2007). https://doi.org/10.1007/s00412-007-0111-y

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