BUD6 Antibody
Description
Overview of Bud6 Protein and Antibody Utility
Bud6 (YLR319C) is a multifunctional protein involved in actin nucleation, formin regulation, and cortical tethering. Antibodies against Bud6 enable researchers to:
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Detect Bud6 expression levels in wild-type and mutant strains
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Localize Bud6 via immunofluorescence microscopy
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Investigate protein-protein interactions (e.g., with formins Bni1/Bnr1)
| Property | Value |
|---|---|
| Molecular Weight | 88.8 kDa |
| Isoelectric Point (pI) | 7.95 |
| Cellular Abundance | ~2,208 molecules/cell |
Functional Domains and Epitope Mapping
Bud6 contains two critical regions for antibody targeting:
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N-terminal domain (residues 1–488): Mediates cortical localization and microtubule interactions .
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C-terminal domain (residues 489–788): Binds formins (Bni1/Bnr1) and actin monomers, essential for nucleation-promoting factor (NPF) activity .
Critical residues identified through mutagenesis :
| Mutation | Functional Defect |
|---|---|
| Bud6-35 | Disrupted Bni1 binding (DAD domain) |
| Bud6-8 | Impaired G-actin recruitment |
| E585S | Reduced Bni1 interaction |
Actin Nucleation and Formin Regulation
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Bud6 enhances Bni1-mediated actin nucleation by recruiting G-actin to the formin’s DAD domain, increasing filament density by 3–5× .
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In bud6Δ cells, actin cable staining is severely diminished, and polarized growth is disrupted .
Cortical Localization and Autoinhibition
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Bud6 anchors the formin For3p to cell tips in Schizosaccharomyces pombe. Deletion of BUD6 causes mislocalization of For3p’s C-terminal fragment .
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Antibody studies show Bud6 counteracts formin autoinhibition by binding near the DAD domain .
Genetic Interactions
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bud6Δ exhibits synthetic lethality with bni1Δ and hypersensitivity to actin-depolymerizing agents (e.g., latrunculin A) .
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Strain LatA Sensitivity (IC₅₀) Wild-type 50 µM Δaip5 15 µM Δbud6 30 µM
Antibody Validation in Key Studies
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Co-immunoprecipitation: Confirmed Bud6-Bni1/Bnr1 interactions in bni1Δbud6-35 and bni1Δbud6-8 mutants .
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TIRF microscopy: Demonstrated Bud6’s role in increasing Bnr1-dependent filament nucleation rates .
Future Directions
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Develop isoform-specific antibodies to distinguish Bud6’s roles in actin assembly vs. microtubule capture.
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Investigate Bud6’s post-translational modifications in response to cellular stress.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Function of BUD6
BUD6 has been implicated in various cellular processes related to actin organization and microtubule dynamics. Here are some key findings:
- Structure of a Bud6/Actin complex reveals a novel wh2-like actin monomer recruitment motif. PMID: 26118535
- Yor304c-a/Bil1 colocalizes with Bud6 and functions in the Bnr1 pathway for actin assembly PMID: 23671312
- A model has been proposed where a Bni1/Bud6 complex with a 2:4 subunit stoichiometry assembles a nucleation seed with Bud6 coordinating up to four actin subunits. PMID: 23161908
- Bud6p and Kip3p play antagonistic roles in controlling the length of microtubules contacting the bud. PMID: 22608510
- Bud6 promotes actin filament nucleation by recruiting actin monomers to Bni1. PMID: 21880892
- Bud6p contributes to MT capture at the cell cortex, independent of Kar9p-mediated MT delivery along actin cables. PMID: 15492045
- Bni1 and Bnr1 are regulated by Bud6. PMID: 15923184
- Bud6p, through its localization at both spindle pole bodies and at the mother-bud neck, provides a regulatory mechanism to prevent excessive oscillations of preanaphase nuclei, thus reducing the likelihood of mitotic delays and nuclear missegregation. PMID: 17416900
- BNI1 and BNR1 contribute to Bud6p-dependent microtubule-cortex interactions by controlling Bud6p localisation. PMID: 18957510
KEGG: sce:YLR319C
STRING: 4932.YLR319C
