PMID:18552200

Citation	Zhang, Y, Feng, S, Chen, F, Chen, H, Wang, J, McCall, C, Xiong, Y and Deng, XW (2008) Arabidopsis DDB1-CUL4 ASSOCIATED FACTOR1 forms a nuclear E3 ubiquitin ligase with DDB1 and CUL4 that is involved in multiple plant developmental processes. Plant Cell 20:1437-55
Abstract	The human DDB1-CUL4 ASSOCIATED FACTOR (DCAF) proteins have been reported to interact directly with UV-DAMAGED DNA BINDING PROTEIN1 (DDB1) through the WDxR motif in their WD40 domain and function as substrate-recognition receptors for CULLIN4-based E3 ubiquitin ligases. Here, we identified and characterized a homolog of human DCAF1/VprBP in Arabidopsis thaliana. Yeast two-hybrid analysis demonstrated the physical interaction between DCAF1 and DDB1 from Arabidopsis, which is likely mediated via the WD40 domain of DCAF1 that contains two WDxR motifs. Moreover, coimmunoprecipitation assays showed that DCAF1 associates with DDB1, RELATED TO UBIQUITIN-modified CUL4, and the COP9 signalosome in vivo but not with CULLIN-ASSOCIATED and NEDDYLATION-DISSOCIATED1, CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1), or the COP10-DET1-DDB1 complex, supporting the existence of a distinct Arabidopsis CUL4 E3 ubiquitin ligase, the CUL4-DDB1-DCAF1 complex. Transient expression of fluorescently tagged DCAF1, DDB1, and CUL4 in onion epidermal cells showed their colocalization in the nucleus, consistent with the notion that the CUL4-DDB1-DCAF1 complex functions as a nuclear E3 ubiquitin ligase. Genetic and phenotypic analysis of two T-DNA insertion mutants of DCAF1 showed that embryonic development of the dcaf1 homozygote is arrested at the globular stage, indicating that DCAF1 is essential for plant embryogenesis. Reducing the levels of DCAF1 leads to diverse developmental defects, implying that DCAF1 might be involved in multiple developmental pathways.
Links	PubMed PMC2483375 Online version:10.1105/tpc.108.058891
Keywords	Amino Acid Motifs; Amino Acid Sequence; Arabidopsis/embryology; Arabidopsis/genetics; Arabidopsis/metabolism; Arabidopsis Proteins/genetics; Arabidopsis Proteins/metabolism; Arabidopsis Proteins/physiology; Cullin Proteins/genetics; Cullin Proteins/metabolism; Cullin Proteins/physiology; DNA, Bacterial/genetics; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; DNA-Binding Proteins/physiology; Gene Expression Regulation, Developmental; Gene Expression Regulation, Plant; Humans; Immunoprecipitation; Molecular Sequence Data; Mutagenesis, Insertional; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Nuclear Proteins/physiology; Onions/genetics; Onions/metabolism; Plants, Genetically Modified/embryology; Plants, Genetically Modified/genetics; Plants, Genetically Modified/metabolism; Protein Binding; Sequence Homology, Amino Acid; Ubiquitin-Conjugating Enzymes/genetics; Ubiquitin-Conjugating Enzymes/metabolism; Ubiquitin-Conjugating Enzymes/physiology
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