We have undertaken a study of the yeast cullin family members Cul3 and Cul8, as little is known about their biochemical and physiological functions. We demonstrate that these cullins are associated in vivo with ubiquitin ligase activity. We show that Cul3 and Cul8 are functionally distinct from Cdc53 and do not interact with ySkp1, suggesting that they target substrates by Skp1- and possibly F-box protein-independent mechanisms. Whereas null mutants of CUL3 appear normal, yeast cells lacking CUL8 have a slower growth rate and are delayed in their progress through anaphase. The anaphase delay phenotype can be complemented by ectopic expression of Cul8 but not by any other yeast or human cullins, nor by a cul8 mutant deficient in binding to RING finger protein Roc1. Deletion of the RAD9 gene suppressed the anaphase delay phenotype of cul8delta, suggesting that loss of Cul8 function may compromise genomic integrity. These results indicate that in addition to the anaphase promoting complex, mitotic progression may involve another E3 ubiquitin ligase mediated by Cul8 protein.
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| Gene/Complex | Qualifier | Gene Ontology Term | Annotation Extension | Evidence | Source | Assigned On |
|---|---|---|---|---|---|---|
| CUL3 | enables | ubiquitin-protein transferase activity | IDA | SGD | 2013-08-07 | |
| RTT101 | enables | ubiquitin-protein transferase activity | IDA | SGD | 2013-08-07 | |
| RTT101 | involved in | regulation of mitotic nuclear division | IMP | SGD | 2013-08-07 | |
| HRT1 | part of | Cul8-RING ubiquitin ligase complex | IDA | SGD | 2013-08-07 | |
| HRT1 | part of | Cul3-RING ubiquitin ligase complex | IDA | SGD | 2013-08-07 |
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| Gene | Phenotype | Experiment Type | Mutant Information | Strain Background | Chemical | Details |
|---|---|---|---|---|---|---|
| RTT101 | budding index: increased | classical genetics | null Allele: rtt101-Δ | S288C | Details: increase in large budded cells from 25.5% in wt to 37.2% in the null | |
| RTT101 | cell cycle progression in mid anaphase: delayed | classical genetics | null Allele: rtt101-Δ | S288C | Details: mutant cells delay with DNA positioned midway in the bud neck (14% vs 0% in wt) | |
| RTT101 | growth in exponential phase: decreased rate | classical genetics | null Allele: rtt101-Δ | S288C | ||
| RTT101 | viable | classical genetics | null Allele: rtt101-Δ | S288C | ||
| CUL3 | viable | classical genetics | null Allele: cul3-Δ | S288C |
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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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| Interactor | Interactor | Assay | Annotation | Action | Modification | |
|---|---|---|---|---|---|---|
| CDC53 | SKP1 | Two-hybrid | manually curated | Bait-Hit | No Modification | |
| CUL3 | HRT1 | Two-hybrid | manually curated | Bait-Hit | No Modification | |
| RTT101 | HRT1 | Two-hybrid | manually curated | Bait-Hit | No Modification | |
| RTT101 | HRT1 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification |