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Dual functions for the ssDNA-binding protein RPA in meiotic recombination
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Dual functions for the ssDNA-binding protein RPA in meiotic recombination
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Journal Article
Dual functions for the ssDNA-binding protein RPA in meiotic recombination
2019
Overview
Meiotic recombination permits exchange of genetic material between homologous chromosomes. The replication protein A (RPA) complex, the predominant ssDNA-binding complex, is required for nearly all aspects of DNA metabolism, but its role in mammalian meiotic recombination remains unknown due to the embryonic lethality of RPA mutant mice. RPA is a heterotrimer of RPA1, RPA2, and RPA3. We find that loss of RPA1, the largest subunit, leads to disappearance of RPA2 and RPA3, resulting in the absence of the RPA complex. Using an inducible germline-specific inactivation strategy, we find that loss of RPA completely abrogates loading of RAD51/DMC1 recombinases to programmed meiotic DNA double strand breaks, thus blocking strand invasion required for chromosome pairing and synapsis. Surprisingly, loading of MEIOB, SPATA22, and ATR to DNA double strand breaks is RPA-independent and does not promote RAD51/DMC1 recruitment in the absence of RPA. Finally, inactivation of RPA reduces crossover formation. Our results demonstrate that RPA plays two distinct roles in meiotic recombination: an essential role in recombinase recruitment at early stages and an important role in promoting crossover formation at later stages.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ Animals
/ Cell Cycle Proteins - metabolism
/ DNA
/ DNA, Single-Stranded - metabolism
/ DNA-Binding Proteins - metabolism
/ Embryos
/ Funding
/ Genes
/ Genetics
/ Genomes
/ Male
/ Males
/ Medicine and Health Sciences
/ Meiosis
/ Methods
/ Mice
/ Mutation
/ Nuclear Proteins - metabolism
/ Proteins
/ Rad51 Recombinase - deficiency
/ Rad51 Recombinase - genetics
/ Rad51 Recombinase - metabolism
/ Replication Protein A - deficiency
/ Replication Protein A - genetics
/ Replication Protein A - metabolism
/ Research and Analysis Methods
/ Yeast
