Dbf4 recruitment by forkhead transcription factors defines an upstream rate-limiting step in determining replication origin firing

Initiation of eukaryotic chromosome replication follows a spatiotemporal program. Current model suggests that replication origins compete for a limited pool of initiation factors. However, it remains to be answered how these limiting factors are preferentially recruited to early origins. Here, we report that Dbf4 is enriched at early origins through its interaction with forkhead transcription factors Fkh1 and Fkh2. This interaction is mediated by Dbf4 C-terminus and was successfully reconstituted in vitro. An interaction defective mutant dbf4?C phenocopies fkh alleles in terms of origin firing. Remarkably, genome-wide replication profiles reveal that the direct fusion of the DNA-binding domain of Fkh1 to Dbf4 restores the Fkh-dependent origin firing, but specifically interferes with the pericentromeric origin activation. Furthermore, Dbf4 directly interacts with Sld3 and promotes the recruitment of downstream limiting factors. These data suggest that Fkh1 targets Dbf4 to a subset of non-centromeric origins to promote early replication, in a manner that is reminiscent to the recruitment of Dbf4 to pericentromeric origins by Ctf19. Overall design: NGS-based analysis of origin activity in HU-arrested S. cerevisiae cells

Identifier
Source https://data.blue-cloud.org/search-details?step=~0120601E1AAB2A0F313CC2633CB6DE2F40494883FC4
Metadata Access https://data.blue-cloud.org/api/collections/0601E1AAB2A0F313CC2633CB6DE2F40494883FC4
Provenance
Instrument Illumina HiSeq 4000; Illumina HiSeq 2000; ILLUMINA
Publisher Blue-Cloud Data Discovery & Access service; ELIXIR-ENA
Contributor IGH
Publication Year 2024
OpenAccess true
Contact blue-cloud-support(at)maris.nl
Representation
Discipline Marine Science
Temporal Point 2018-01-24T00:00:00Z