Reference: Kile AC and Koepp DM (2010) Activation of the S-phase checkpoint inhibits degradation of the F-box protein Dia2. Mol Cell Biol 30(1):160-71

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Abstract


A stable genome is critical to cell viability and proliferation. During DNA replication, the S-phase checkpoint pathway responds to replication stress. In budding yeast, the chromatin-bound F-box protein Dia2 is required to maintain genomic stability and may help replication complexes overcome sites of damaged DNA and natural fragile regions. SCF (Skp1/Cul1/F-box protein) complexes are modular ubiquitin ligases. We show here that Dia2 is itself targeted for ubiquitin-mediated proteolysis and that activation of the S-phase checkpoint pathway inhibits Dia2 protein degradation. S-phase checkpoint mutants fail to stabilize Dia2 in response to replication stress. Deletion of DIA2 from these checkpoint mutants exacerbates their sensitivity to hydroxyurea, suggesting that stabilization of Dia2 contributes to the replication stress response. Unlike the case for other F-box proteins, deletion of the F-box domain in Dia2 does not stabilize the protein. Rather, an N-terminal domain that is also required for nuclear localization is necessary for degradation. When a strong nuclear localization signal (NLS) is added to dia2 mutants lacking this domain, the Dia2 protein is both stable and nuclear. Together, our results suggest that Dia2 protein turnover does not involve an autocatalytic mechanism and that Dia2 proteolysis is inhibited by activation of the replication stress response.

Reference Type
Journal Article | Research Support, N.I.H., Extramural | Research Support, Non-U.S. Gov't
Authors
Kile AC, Koepp DM
Primary Lit For
SKP1 | CDC53 | DIA2 | SCF-Dia2 ubiquitin ligase complex
Additional Lit For
CHK1 | RAD24 | MRC1 | RAD17 | cdc53-1 | skp1-11

Gene Ontology Annotations 1 entry for 1 gene


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Gene/ComplexQualifierGene Ontology TermAnnotation ExtensionEvidenceSourceAssigned On
DIA2located innucleusIDASGD2013-08-07
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Phenotype Annotations 3 entries for 3 genes


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GenePhenotypeExperiment TypeMutant InformationStrain BackgroundChemicalDetails
SKP1protein/peptide accumulation: decreased
Reporter: Dia2-Myc
classical geneticsconditional
Allele: skp1-11

G160E, R167K

Other100 ug/ml cycloheximideTemperature: elevated temperature, 37 °C
CDC53protein/peptide accumulation: decreased
Reporter: Dia2-Myc
classical geneticsconditional
Allele: cdc53-1
Other100 ug/ml cycloheximideTemperature: elevated temperature, 37 °C
DIA2vegetative growth: decreased
classical geneticsoverexpressionOtherDetails: mild defect; growth defect severe in the presence of 0.005% methylmethanesulfonate
Showing 1 to 3 of 3 entries

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 1 entry for 2 genes

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InteractorInteractorAssayAnnotationActionModification
SKP1DIA2Affinity Capture-Westernmanually curatedBait-HitNo Modification
Showing 1 to 1 of 1 entries