Unknown

Dataset Information

0

DNA replication checkpoint signaling depends on a Rad53-Dbf4 N-terminal interaction in Saccharomyces cerevisiae.


ABSTRACT: Dbf4-dependent kinase (DDK) and cyclin-dependent kinase (CDK) are essential to initiate DNA replication at individual origins. During replication stress, the S-phase checkpoint inhibits the DDK- and CDK-dependent activation of late replication origins. Rad53 kinase is a central effector of the replication checkpoint and both binds to and phosphorylates Dbf4 to prevent late-origin firing. The molecular basis for the Rad53-Dbf4 physical interaction is not clear but occurs through the Dbf4 N terminus. Here we found that both Rad53 FHA1 and FHA2 domains, which specifically recognize phospho-threonine (pT), interacted with Dbf4 through an N-terminal sequence and an adjacent BRCT domain. Purified Rad53 FHA1 domain (but not FHA2) bound to a pT Dbf4 peptide in vitro, suggesting a possible phospho-threonine-dependent interaction between FHA1 and Dbf4. The Dbf4-Rad53 interaction is governed by multiple contacts that are separable from the Cdc5- and Msa1-binding sites in the Dbf4 N terminus. Importantly, abrogation of the Rad53-Dbf4 physical interaction blocked Dbf4 phosphorylation and allowed late-origin firing during replication checkpoint activation. This indicated that Rad53 must stably bind to Dbf4 to regulate its activity.

SUBMITTER: Chen YC 

PROVIDER: S-EPMC3664849 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

DNA replication checkpoint signaling depends on a Rad53-Dbf4 N-terminal interaction in Saccharomyces cerevisiae.

Chen Ying-Chou YC   Kenworthy Jessica J   Gabrielse Carrie C   Hänni Christine C   Zegerman Philip P   Weinreich Michael M  

Genetics 20130405 2


Dbf4-dependent kinase (DDK) and cyclin-dependent kinase (CDK) are essential to initiate DNA replication at individual origins. During replication stress, the S-phase checkpoint inhibits the DDK- and CDK-dependent activation of late replication origins. Rad53 kinase is a central effector of the replication checkpoint and both binds to and phosphorylates Dbf4 to prevent late-origin firing. The molecular basis for the Rad53-Dbf4 physical interaction is not clear but occurs through the Dbf4 N termin  ...[more]

Similar Datasets

| S-EPMC3268399 | biostudies-literature
| S-EPMC1142443 | biostudies-literature
| S-EPMC3834268 | biostudies-literature
| S-EPMC4218987 | biostudies-literature
| S-EPMC2064673 | biostudies-literature
| S-EPMC3024743 | biostudies-literature
| S-EPMC3908393 | biostudies-literature
| S-EPMC8387023 | biostudies-literature
| S-EPMC2948544 | biostudies-literature
| S-EPMC2795593 | biostudies-literature