Rad53, the ortholog of mammalian Chk2, is a major DNA damage checkpoint effector kinase in Saccharomyces cerevisiae. Despite extensive studies on the genetic requirements for Rad53 activation and its linkage downstream to checkpoint responses, the mechanism of Rad53 activation is not completely understood. Rad53-dependent signal amplification is thought to be a primary force that accelerates checkpoint signal transduction processes in response to DNA damage. Rad53 forms oligomers upon DNA damage in vivo. It is not clear how oligomer formation affects Rad53 activation and what is the mechanism of Rad53 oligomerization. Here, we monitor Rad53 oligomer assembly and disassembly in vitro. These processes are ATP-dependent and are regulated through phosphorylation. Mutations in FHA or SCD domains of RAD53 compromise intermolecular autophosphorylation activity and these domains are indispensable for Rad53 oligomerization. The mediator Rad9 is not necessary for Rad53 oligomerization. Rad53 kinase activity is required for disassembly of Rad53 oligomers in vivo after DNA damage. Moreover, induced oligomerization of Rad53 efficiently activates Rad53 in the absence of Mec1 in vivo. The results support the conclusions that Rad53/Chk2 homo-oligomerization is an evolutionarily conserved mechanism that drives Rad53/Chk2 activation and promotes signal amplification in DNA damage responses.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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