Background: Correct positioning of the mitotic spindle relative to the cleavage apparatus is crucial for successful mitosis. In budding yeast, the Adenomatous Polyposis Coli-related protein Kar9, yeast EB1, and Myo2, a type V myosin, mediate positioning of the mitotic spindle close to the septin-anchored cleavage apparatus at the bud neck.
Results: We find that Kar9 is ubiquitylated and degraded by the proteasome. Ubiquitylation requires the ubiquitin-conjugating enzymes Ubc1 and Ubc4 and phosphorylation of Kar9 by yeast Cdk1. Importantly, Kar9 ubiquitylation and degradation depend on an intact cleavage apparatus. Kar9 is stabilized in septin mutant cells or cells lacking the bud neck formin Bnr1, but not in the bud formin Bni1 or the actomyosin ring. Transport of Kar9 to the bud neck by Myo2 is also required for Kar9 degradation. Abrogation of Kar9 phosphorylation and ubiquitylation increases interactions of astral microtubules (aMTs) with the bud neck and causes spindle mispositioning. Photoconversion experiments showed that Kar9 association with aMTs is stable.
Conclusions: We propose that ubiquitylation controls interactions of aMTs with the cleavage apparatus through localized disassembly of Kar9 complexes.
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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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Evidence ID | Analyze ID | File | Description |
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