FUS3 Antibody
Description
Applications of FUS3 Antibodies
FUS3 antibodies are utilized across multiple research domains:
Table 1: Key Applications of FUS3 Antibodies
Fungal Pathogenesis and Aflatoxin Biosynthesis
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In Aspergillus flavus, Fus3 regulates mycelial growth, conidiation, and aflatoxin B1 (AFB1) production. Deletion of fus3 reduces AFB1 by 80% due to decreased acetyl-CoA and malonyl-CoA levels, critical substrates for toxin synthesis .
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Transcriptome and phosphoproteome analyses revealed Fus3 modulates genes involved in lipid metabolism (e.g., accA), linking kinase activity to secondary metabolite production .
Yeast Mating and Signal Transduction
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Fus3 orchestrates mating responses in S. cerevisiae by phosphorylating Far1 (a cyclin-dependent kinase inhibitor) and repressing G1/S cyclin genes .
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Oscillatory phosphorylation of Fus3 drives pulsatile gene expression (e.g., FUS1), with peak activity correlating with transcriptional bursts .
Plant Development
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In Arabidopsis thaliana, FUS3 directly binds promoters of embryo-specific genes (e.g., ABI3, LEC1), regulating seed maturation and dormancy .
Future Directions and Therapeutic Potential
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Engineered Fusion Proteins: Antibody-Fus3 fusions could enhance targeted therapies, as seen in 3E10-based platforms delivering enzymes like myotubularin to treat muscular dystrophies .
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Agricultural Biotechnology: Modulating Fus3 in crops may reduce mycotoxin contamination by disrupting aflatoxin pathways .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- AIP2 targets FUS3 for degradation and plays a role in cotyledon development and flowering time in Arabidopsis. PMID: 28369580
- FUS3 phosphorylation and SnRK1 are essential for embryogenesis and integrating environmental cues to ensure progeny survival. PMID: 28922765
- LEAFY COTYLEDON2 (LEC2) interacts with FUS3. These homologous B3 transcription factors bind to the auxin biosynthesis gene YUCCA4 (YUC4) and synergistically activate its transcription during lateral roots formation. PMID: 27878992
- TT2 binds directly to the regulatory region of FUSCA3 and mediates the expression of numerous genes involved in the fatty acid biosynthesis pathway. PMID: 24397827
- Analysis of direct FUS3 targets indicates that it primarily or exclusively acts as a transcriptional activator. PMID: 23314941
- Activation of FUS3 after germination reduces the expression of genes involved in ethylene biosynthesis and response, while a loss-of-function fus3 mutant exhibits phenotypes consistent with increased ethylene signaling. PMID: 22348746
- FUS3 contributes to delaying seed germination at high temperatures. PMID: 22279962
- The genes positively regulated by FUS3 extend beyond known seed maturation-related genes and encompass those involved in secondary metabolite production and primary metabolism. PMID: 21045071
- The C-terminal domain of FUS3 is crucial for its normal function and sensitivity to abscisic acid and gibberellic acid. It negatively regulates mRNA and protein levels. PMID: 20663088
- Research has demonstrated that gibberellin (GA) hormone biosynthesis is regulated by the LEC2 and FUS3 pathways. PMID: 15516508
- At2S3 activation by FUS3 is rapid, while CRC induction by FUS3 with abscissic acid [ABA] and by ABA followed by FUS3 presence is slower, suggesting an indirect mechanism involving the synthesis of intermediate regulatory factors. PMID: 15695463
- Double (lec1 lec2, lec1 fus3, lec2 fus3) and triple (fus3 lec1 lec2) mutants exhibit total repression of embryogenic potential. PMID: 16034595
- Studies show that significant differences in histone modifications at the phas promoter are mediated by FUS3 and PvALF, indicating that they function through distinct epigenetic mechanisms. PMID: 18038114
- FUS3 function is limited to acquiring embryo-dependent seed dormancy, determining cotyledonary cell identity, and synthesizing and accumulating storage compounds. PMID: 18343361
Q&A
FAQs for FUS3 Antibody in Academic Research
FUS3 antibodies are critical tools for studying MAPK signaling pathways across organisms. Below are FAQs addressing methodological challenges and advanced research applications, supported by experimental evidence from peer-reviewed studies.
What experimental designs optimize FUS3 antibody use in immunofluorescence?
Advanced Protocol:
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Fixation: Paraformaldehyde (4%) preserves FUS3 punctate cytoplasmic and nuclear localization .
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Signal Enhancement: Pre-treat cells with pheromones (e.g., α-factor in yeast) to increase nuclear FUS3 localization .
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Controls: Co-stain with nuclear markers (e.g., DAPI) to validate subcellular distribution patterns.
Key Finding:
"Fus3 localizes in punctate spots throughout the cytoplasm and nucleus, with enhanced nuclear localization post-pheromone stimulation" .
How to resolve contradictions in FUS3 antibody performance across species?
Analysis Framework:
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Epitope Conservation: Compare FUS3 protein sequences (e.g., Arabidopsis vs. fungi) to identify cross-reactive epitopes .
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Functional Redundancy: Assess compensatory mechanisms (e.g., Kss1 in yeast) that may mask antibody detection failures .
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Technical Validation: Perform reciprocal co-IP or CRISPR-Cas9 tagging to confirm antibody reliability .
How to design co-IP experiments for studying FUS3-MAPK interactions?
Step-by-Step Workflow:
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Complex Stabilization: Use ste50, ste11, and ste7 mutants to isolate FUS3 interaction dependencies .
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Gradient Centrifugation: Separate FUS3 complexes by size (e.g., 350–500 kDa complexes show highest kinase activity) .
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Phosphorylation Mapping: Combine co-IP with mass spectrometry to identify interacting partners and phosphorylation sites .
Critical Data:
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Trimeric Ste11-Ste7-Fus3 interactions are essential for phosphorylation signal transduction .
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Monomeric FUS3 exhibits negligible kinase activity compared to complex-bound forms .
What methodologies analyze FUS3 phosphorylation dynamics during fungal development?
Integrated Approach:
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Phosphoproteomics: Quantify phosphorylation changes using MS-based assays (e.g., 11,703 phosphorylation sites identified in A. flavus) .
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Transcriptome Correlation: Cross-reference phosphoproteomic data with RNA-seq results (e.g., 4,437 DEGs in Δfus3) to identify regulatory hubs .
Example Workflow:
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Induce fungal development (e.g., sclerotia formation in WKM media).
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Extract proteins at critical growth phases (mycelia, conidia, sclerotia).
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Perform anti-FUS3 immunoprecipitation followed by LC-MS/MS.
How to address discrepancies in FUS3’s role in secondary metabolism vs. development?
Hypothesis Testing:
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Substrate Limitation: In Δfus3, reduced acetyl-CoA/malonyl-CoA levels (↓30–50%) explain aflatoxin reduction despite upregulated biosynthetic genes .
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Kinase Targets: Validate direct targets (e.g., AccA in acetyl-CoA biosynthesis) via site-directed mutagenesis and enzymatic assays .
Contradiction Resolution Table:
