Rad51–Rad52 Mediated Maintenance of Centromeric Chromatin in Candida albicans

摘要: Specification of the centromere location in most eukaryotes is not solely dependent on DNA sequence. However, non-genetic determinants identity are clearly defined. While multiple mechanisms, individually or concert, may specify centromeres epigenetically, studies this area focused a universal factor, centromere-specific histone H3 variant CENP-A, often considered as epigenetic determinant identity. In spite variable timing its loading at across species, replication coupled early S phase deposition CENP-A found yeast centromeres. Centromeres earliest replicating chromosomal regions pathogenic budding Candida albicans. Using 2-dimensional agarose gel electrophoresis assay, we identify origins (ORI7-LI and ORI7-RI) proximal to an (CEN7) C. We show that forks stall CEN7 kinetochore manner fork stalling reduced absence homologous recombination (HR) proteins Rad51 Rad52. Deletion ORI7-RI causes significant reduction stalled signal increased loss rate altered chromosome 7. The HR proteins, Rad52, have been shown play role restart. Confocal microscopy shows declustered kinetochores rad51 rad52 mutants, which evidence disintegrity. CENP-ACaCse4 levels centromeres, determined by chromatin immunoprecipitation (ChIP) experiments, Rad51/Rad52 resulting disruption structure. Moreover, western blot analysis reveals delocalized molecules mutants degrade similar fashion other described before. Finally, co-immunoprecipitation assays indicate Rad52 physically interact with vivo. Thus, epigenetically maintain functioning regulating programmed sites