Reference: Winter DC, et al. (1999) Genetic dissection of the budding yeast Arp2/3 complex: a comparison of the in vivo and structural roles of individual subunits. Proc Natl Acad Sci U S A 96(13):7288-93

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Abstract


In previous work, we identified the yeast Arp2/3 complex, which localizes to cortical actin patches and is required for their motility and integrity in vivo. This complex contains proteins homologous to each subunit of the Acanthamoeba and human Arp2/3 complex except for a 40-kDa subunit (p40), which was missing from the purified yeast complex. Here, we demonstrate by using immunoprecipitation and gel-filtration analysis that Arc40p, the homolog of p40 identified from the yeast genome database, associates with the yeast Arp2/3 complex. We have carried out gene disruptions of each subunit of the yeast Arp2/3 complex to study each subunit's role in the function of the complex. Surprisingly, we find that only ARC40 is fully essential for cell viability. Strains lacking each of the other subunits exhibit varying degrees of defects in cell growth and viability and in assembly and polarization of cortical actin patches. We have also examined each subunit's role in maintaining the structural integrity of the Arp2/3 complex. Arp2p, Arp3p, and Arc40p fall into the monomer pool in Deltaarc19 and Deltaarc35 cells, suggesting that Arc19p and Arc35p are the central scaffolding components of the complex. Arp2p and Arp3p do not have major roles in maintaining complex integrity, and Arc15p is required for association of Arp2p and Arc40p, but not other subunits, with the complex. These results provide evidence that each subunit contributes differently to the assembly and function of the Arp2/3 complex.

Reference Type
Comparative Study | Journal Article | Research Support, U.S. Gov't, P.H.S.
Authors
Winter DC, Choe EY, Li R
Primary Lit For
ARP3 | ARC15 | ARC19 | ARP2 | ARC40 | ARC18 | ARC35 | Actin-related protein 2/3 complex
Additional Lit For
ACT1 | arc40-Δ

Gene Ontology Annotations


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Gene/Complex Qualifier Gene Ontology Term Aspect Annotation Extension Evidence Method Source Assigned On Reference

Phenotype Annotations 21 entries for 6 genes


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GenePhenotypeExperiment TypeMutant InformationStrain BackgroundChemicalDetails
ARC35actin cytoskeleton morphology: abnormal
classical geneticsnull
Allele: arc35-Δ
S288CDetails: aberrant structures form in place of actin patches
ARC19actin cytoskeleton morphology: abnormal
classical geneticsnull
Allele: arc19-Δ
S288CDetails: aberrant structures form in place of actin patches
ARC40actin cytoskeleton morphology: abnormal
classical geneticsnull
Allele: arc40-Δ
S288CDetails: aberrant structures form in place of actin patches
ARC15actin cytoskeleton morphology: abnormal
classical geneticsnull
Allele: arc15-Δ
S288CDetails: aberrant structures form in place of actin patches
ARP2actin cytoskeleton morphology: abnormal
classical geneticsnull
Allele: arp2-Δ
S288CDetails: aberrant structures form in place of actin patches
ARP3actin cytoskeleton morphology: abnormal
classical geneticsnull
Allele: arp3-Δ
S288CDetails: aberrant structures form in place of actin patches
ARC35growth in exponential phase: decreased rate
classical geneticsnull
Allele: arc35-Δ
S288CMedia: YPD
Details: doubling time is 7-9 hours, compared to 2-3 hours for wild type cells
ARC19growth in exponential phase: decreased rate
classical geneticsnull
Allele: arc19-Δ
S288CMedia: YPD
Details: doubling time is 7-9 hours, compared to 2-3 hours for wild type cells
ARC40growth in exponential phase: decreased rate
classical geneticsnull
Allele: arc40-Δ
S288CMedia: YPD
Details: doubling time is 7-9 hours, compared to 2-3 hours for wild type cells
ARC15growth in exponential phase: decreased rate
classical geneticsnull
Allele: arc15-Δ
S288CMedia: YPD
Details: doubling time is 7-9 hours, compared to 2-3 hours for wild type cells
Showing 1 to 10 of 21 entries

Interaction Annotations


Genetic Interactions

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

Physical Interactions 10 entries for 7 genes

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InteractorInteractorAssayAnnotationActionModification
ARC15ARP3Affinity Capture-Westernmanually curatedHit-BaitNo Modification
ARC19ARP3Affinity Capture-Westernmanually curatedHit-BaitNo Modification
ARC40ARP2Affinity Capture-Westernmanually curatedBait-HitNo Modification
ARC40ARP3Affinity Capture-Westernmanually curatedBait-HitNo Modification
ARC40ARP3Affinity Capture-Westernmanually curatedHit-BaitNo Modification
ARC40ARP3Co-purificationmanually curatedHit-BaitNo Modification
ARP2ARP3Affinity Capture-Westernmanually curatedHit-BaitNo Modification
ARP2ARP3Co-purificationmanually curatedHit-BaitNo Modification
ARP3ARC18Affinity Capture-Westernmanually curatedBait-HitNo Modification
ARP3ARC35Affinity Capture-Westernmanually curatedBait-HitNo Modification
Showing 1 to 10 of 10 entries