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Both conserved and non-conserved regions of Spo11 are essential for meiotic recombination initiation in yeast

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Abstract

DNA double-strand breaks (DSBs) are the initiators of most meiotic recombination events. In Saccharomyces cerevisiae, at least ten genes are necessary for meiotic DSB formation. However, the molecular roles of these proteins are not clearly understood. The meiosis-specific Spo11 protein, which shows sequence similarity with a subunit of an archaeal topoisomerase, is believed to catalyze the meiotic DSB formation. Spo11 is also required for induction of meiotic DSBs at long inverted repeats and at large trinucleotide repeat tracts. Here we report the isolation and characterization of temperature-sensitive spo11-mutant alleles to better understand how Spo11 functions, and how meiotic DSBs are generated at various recombination hotspots. Analysis of mutation sites of isolated spo11-mutant alleles indicated that both N-terminal and C-terminal non-conserved residues of Spo11 are essential for the protein’s function, possibly for interaction with other meiotic DSB enzymes. Several of the mutation sites within the conserved region are predicted to lie on the surface of the protein, suggesting that this region is required for activation of the meiotic initiation complex via protein-protein interaction. In addition to the conditional mutants, we isolated partially recombination-defective mutants; analysis of one of these mutants indicated that Ski8, as observed previously, interacts with Spo11 via the latter’s C-terminal residues.

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Acknowledgments

We thank Kathy Atcheson and Scott Keeney for generously providing plasmids used in this study. We also thank the Wadsworth Center Molecular Genetics Core Facility for sequencing the spo11-mutant alleles. Janice Pata was supported by the new investigator start-up award from Health Research Inc., Albany, NY, USA.

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Author notes

  1. Dilip K. Nag

    Present address: Laboratory of Cell Regulation, Wadsworth Center, P. O. Box 509, Albany, NY, 12201-0509, USA

Authors and Affiliations

  1. Laboratory of Developmental Genetics, Wadsworth Center, Center for Medical Sciences, 150 New Scotland Avenue, Albany, NY, 12208, USA

    Dilip K. Nag, Manuela Sironi, David R. Flood & Ashley M. Hart

  2. Laboratory of Molecular Diagnostics, Wadsworth Center, Center for Medical Sciences, 150 New Scotland Avenue, Albany, NY, 12208, USA

    Janice D. Pata

  3. Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, NY, 12201, USA

    Dilip K. Nag & Janice D. Pata

  4. Scientific Institute IRCCS E. Medea, Via Don Luigi Monza 20, 23842, Bosisio Parini (LC), Italy

    Manuela Sironi

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  1. Dilip K. Nag
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  2. Janice D. Pata
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Correspondence to Dilip K. Nag.

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Communicated by A. Aguilera

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Nag, D.K., Pata, J.D., Sironi, M. et al. Both conserved and non-conserved regions of Spo11 are essential for meiotic recombination initiation in yeast. Mol Genet Genomics 276, 313–321 (2006). https://doi.org/10.1007/s00438-006-0143-7

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  • DOI: https://doi.org/10.1007/s00438-006-0143-7

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