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PMID: 19763177 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A high throughput genetic screen identifies new early meiotic recombination functions in Arabidopsis thaliana.

PLoS genetics ·Vol. 5 ·No. 9 ·2009-09-00 ·Pages e1000654

De Muyt A, Pereira L, Vezon D, Chelysheva L, Gendrot G, Chambon A, Lainé-Choinard S, Pelletier G, Mercier R, Nogué F, Grelon M

Abstract

Meiotic recombination is initiated by the formation of numerous DNA double-strand breaks (DSBs) catalysed by the widely conserved Spo11 protein. In Saccharomyces cerevisiae, Spo11 requires nine other proteins for meiotic DSB formation; however, unlike Spo11, few of these are conserved across kingdoms. In order to investigate this recombination step in higher eukaryotes, we took advantage of a high-throughput meiotic mutant screen carried out in the model plant Arabidopsis thaliana. A collection of 55,000 mutant lines was screened, and spo11-like mutations, characterised by a drastic decrease in chiasma formation at metaphase I associated with an absence of synapsis at prophase, were selected. This screen led to the identification of two populations of mutants classified according to their recombination defects: mutants that repair meiotic DSBs using the sister chromatid such as Atdmc1 or mutants that are unable to make DSBs like Atspo11-1. We found that in Arabidopsis thaliana at least four proteins are necessary for driving meiotic DSB repair via the homologous chromosomes. These include the previously characterised DMC1 and the Hop1-related ASY1 proteins, but also the meiotic specific cyclin SDS as well as the Hop2 Arabidopsis homologue AHP2. Analysing the mutants defective in DSB formation, we identified the previously characterised AtSPO11-1, AtSPO11-2, and AtPRD1 as well as two new genes, AtPRD2 and AtPRD3. Our data thus increase the number of proteins necessary for DSB formation in Arabidopsis thaliana to five. Unlike SPO11 and (to a minor extent) PRD1, these two new proteins are poorly conserved among species, suggesting that the DSB formation mechanism, but not its regulation, is conserved among eukaryotes.

MeSH Terms
Alleles Amino Acid Sequence Arabidopsis/cytology,enzymology,genetics Arabidopsis Proteins/chemistry,metabolism Chromosome Pairing/genetics Chromosomes, Plant/genetics Cyclins/chemistry,metabolism DNA Breaks, Double-Stranded Exons/genetics Genes, Plant High-Throughput Screening Assays/methods Introns/genetics Meiosis/genetics Molecular Sequence Data Mutation/genetics Protein Transport Recombinases/metabolism Recombination, Genetic Sequence Alignment
Chemicals
Arabidopsis Proteins Cyclins Recombinases cyclin SDS protein, Arabidopsis
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
De Muyt Arnaud
INRA de Versailles, Institut Jean-Pierre Bourgin, Station de Génétique et d'Amélioration des Plantes UR-254, Versailles, France.
Pereira Lucie
Vezon Daniel
Chelysheva Liudmila
Gendrot Ghislaine
Chambon Aurélie
Lainé-Choinard Sandrine
Pelletier Georges
Mercier Raphaël
Nogué Fabien
Grelon Mathilde
Conflict of Interest

The authors have declared that no competing interests exist.

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Article Info
Journal
PLoS genetics
Abbr.
PLoS Genet
ISSN
1553-7404
DOI
10.1371/journal.pgen.1000654
Published
2009-09-00
Epub
2009-00-18
Pages
e1000654
Language
English
Region
United States
NLM ID
101239074
PMCID
PMC2735182
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pgen.1000654 PMC Full Text View journal details
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