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Rad51-Rad52 Mediated Maintenance of Centromeric Chromatin in Candida albicans

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dc.contributor.author Mitra, Sreyoshi
dc.contributor.author Gomez-Raja, Jonathan
dc.contributor.author Larriba, German
dc.contributor.author Dubey, Dharani Dhar
dc.contributor.author Sanyal, Kaustuv
dc.date.accessioned 2017-02-21T07:11:27Z
dc.date.available 2017-02-21T07:11:27Z
dc.date.issued 2014
dc.identifier.citation Mitra, S; Gomez-Raja, J; Larriba, G; Dubey, DD; Sanyal, K, Rad51-Rad52 Mediated Maintenance of Centromeric Chromatin in Candida albicans. PLoS Genetics 2014, 10 (4), e1004344 http://dx.doi.org/10.1371/journal.pgen.1004344 en_US
dc.identifier.citation PLoS Genetics en_US
dc.identifier.citation 10 en_US
dc.identifier.citation 4 en_US
dc.identifier.issn 1553-7390
dc.identifier.uri https://libjncir.jncasr.ac.in/xmlui/10572/2476
dc.description Open Access en_US
dc.description.abstract Specification of the centromere location in most eukaryotes is not solely dependent on the DNA sequence. However, the non-genetic determinants of centromere identity are not clearly defined. While multiple mechanisms, individually or in concert, may specify centromeres epigenetically, most studies in this area are focused on a universal factor, a centromere-specific histone H3 variant CENP-A, often considered as the epigenetic determinant of centromere identity. In spite of variable timing of its loading at centromeres across species, a replication coupled early S phase deposition of CENP-A is found in most yeast centromeres. Centromeres are the earliest replicating chromosomal regions in a pathogenic budding yeast Candida albicans. Using a 2-dimensional agarose gel electrophoresis assay, we identify replication origins (ORI7-LI and ORI7-RI) proximal to an early replicating centromere (CEN7) in C. albicans. We show that the replication forks stall at CEN7 in a kinetochore dependent manner and fork stalling is reduced in the absence of the homologous recombination (HR) proteins Rad51 and Rad52. Deletion of ORI7-RI causes a significant reduction in the stalled fork signal and an increased loss rate of the altered chromosome 7. The HR proteins, Rad51 and Rad52, have been shown to play a role in fork restart. Confocal microscopy shows declustered kinetochores in rad51 and rad52 mutants, which are evidence of kinetochore disintegrity. CENP-A(CACSe4) levels at centromeres, as determined by chromatin immunoprecipitation (ChIP) experiments, are reduced in absence of Rad51/Rad52 resulting in disruption of the kinetochore structure. Moreover, western blot analysis reveals that delocalized CENP-A molecules in HR mutants degrade in a similar fashion as in other kinetochore mutants described before. Finally, co-immunoprecipitation assays indicate that Rad51 and Rad52 physically interact with CENP-A(CACSe4) in vivo. Thus, the HR proteins Rad51 and Rad52 epigenetically maintain centromere functioning by regulating CENP-A(CACSe4) levels at the programmed stall sites of early replicating centromeres. en_US
dc.description.uri 1553-7404 en_US
dc.description.uri http://dx.doi.org/10.1371/journal.pgen.1004344 en_US
dc.language.iso English en_US
dc.publisher Public Library of Science en_US
dc.rights @Public Library of Science, 2014 en_US
dc.subject Genetics & Heredity en_US
dc.subject Stalled Replication Forks en_US
dc.subject Neighbor-Joining Method en_US
dc.subject Mating-Type Locus en_US
dc.subject Histone Cenp-A en_US
dc.subject DNA-Replication en_US
dc.subject Saccharomyces-Cerevisiae en_US
dc.subject Schizosaccharomyces-Pombe en_US
dc.subject Microtubule Interaction en_US
dc.subject Gene Conversion en_US
dc.subject Budding Yeast en_US
dc.title Rad51-Rad52 Mediated Maintenance of Centromeric Chromatin in Candida albicans en_US
dc.type Article en_US


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