Genomics

Dataset Information

0

Genome-wide EMSA (gEMSA) of ORC and yeast genomic DNA


ABSTRACT: Eukaryotic DNA replication origins are selected in G1-phase when the origin recognition complex (ORC) binds chromosomal DNA, triggering a series of molecular events that culminate in the initiation of DNA replication (a.k.a. origin firing) during S-phase. Each chromosome requires multiple origins for its duplication, and each origin fires at a characteristic time during S-phase, creating a cell-type specific genome replication pattern with relevance to differentiation, genome stability and evolution. It is unclear whether ORC-origin interactions are relevant to the regulation of origin activation time. Here we applied a novel genome-wide strategy to classify origins in the model eukaryote Saccharomyces cerevisiae based on the types of molecular interactions used for ORC-origin binding. Specifically, origins were classified as DNA-dependent when the strength of ORC-origin binding in vivo could be explained by the affinity of ORC for origin DNA in vitro, and, conversely, as ‘chromatin-dependent’ when the ORC-DNA interaction in vitro was insufficient to explain the strength of ORC-origin binding in vivo. The two classes of origins were distinct in terms of local nucleosome architecture and dependence on origin-flanking sequences in plasmid replication assays, consistent with local features of chromatin promoting ORC binding at ‘chromatin-dependent’ origins. Importantly, origins that fired in late S-phase, but not those that fired in early S-phase, showed a significant dependence on the ORC-origin DNA interaction for ORC binding in vivo. Therefore we demonstrate for the first time a connection between ORC-origin binding mechanisms and the regulation of origin activation time at a genome-wide level.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE48440 | GEO | 2013/09/30

SECONDARY ACCESSION(S): PRJNA210146

REPOSITORIES: GEO

Dataset's files

Source:
Action DRS
Other
Items per page:
1 - 1 of 1

Similar Datasets

2013-09-30 | E-GEOD-48440 | biostudies-arrayexpress
2010-03-19 | E-GEOD-16926 | biostudies-arrayexpress
2010-03-19 | GSE16926 | GEO
2009-03-06 | GSE14864 | GEO
2023-03-11 | PXD032977 | Pride
2024-03-18 | GSE249817 | GEO
2022-02-28 | PXD031033 | Pride
2014-02-13 | GSE54923 | GEO
2014-02-13 | E-GEOD-54923 | biostudies-arrayexpress
2009-11-07 | GSE18895 | GEO