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PMID:19153602

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Citation

Osman, K, Sanchez-Moran, E, Mann, SC, Jones, GH and Franklin, FC (2009) Replication protein A (AtRPA1a) is required for class I crossover formation but is dispensable for meiotic DNA break repair. EMBO J. 28:394-404

Abstract

Replication protein A (RPA) is involved in many aspects of DNA metabolism including meiotic recombination. Many species possess a single RPA1 gene but Arabidopsis possesses five RPA1 paralogues. This feature has enabled us to gain further insight into the meiotic role of RPA1. Proteomic analysis implicated one of the AtRPA1 family (AtRPA1a) in meiosis. Immunofluorescence studies confirmed that AtRPA1a is associated with meiotic chromosomes from leptotene through to early pachytene. Analysis of an Atrpa1a mutant revealed that AtRPA1a is not essential at early stages in the recombination pathway. DNA double-strand breaks are repaired in Atrpa1a, but the mutant is defective in the formation of crossovers, exhibiting a 60% reduction in chiasma frequency. Consistent with this, localization of recombination proteins AtRAD51 and AtMSH4 appears normal, whereas the numbers of AtMLH1 and AtMLH3 foci at pachytene are significantly reduced. This suggests that the defect in Atrpa1a is manifested at the stage of second-end capture. Analysis of Atrpa1a/Atmsh4 and Atrpa1a/Atmlh3 double mutants indicates that loss of AtRPA1a predominantly affects the formation of class I, interference-dependent crossovers.

Links

PubMed PMC2646153 Online version:10.1038/emboj.2008.295

Keywords

Arabidopsis; Arabidopsis Proteins/metabolism; Crossing Over, Genetic; DNA Damage; DNA Repair; Genetic Complementation Test; Meiosis; Microscopy, Fluorescence; Models, Biological; Models, Genetic; Mutation; Proteomics/methods; Recombination, Genetic; Replication Protein A/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ARATH:RFA1A

located_in

GO:0000785: chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ARATH:RFA1A

acts_upstream_of_or_within

GO:0007131: reciprocal meiotic recombination

ECO:0000316: genetic interaction evidence used in manual assertion

AGI_LocusCode:AT4G17380

P

Seeded From UniProt

complete

ARATH:RFA1A

acts_upstream_of_or_within

GO:0007131: reciprocal meiotic recombination

ECO:0000316: genetic interaction evidence used in manual assertion

AGI_LocusCode:AT4G35520

P

Seeded From UniProt

complete

ARATH:RFA1A

acts_upstream_of_or_within

GO:0048232: male gamete generation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:RFA1A

acts_upstream_of_or_within

GO:0051026: chiasma assembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:RFA1A

acts_upstream_of_or_within

GO:0009555: pollen development

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:RFA1A

acts_upstream_of_or_within

GO:0007141: male meiosis I

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:MLH1

located_in

GO:0000785: chromatin

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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