Genetic analysis of CAND1-CUL1 interactions in Arabidopsis supports a role for CAND1-mediated cycling of the SCFTIR1 complex.
SKP1-Cullin1-F-box protein (SCF) ubiquitin-ligases regulate numerous aspects of eukaryotic growth and development. Cullin-Associated and Neddylation-Dissociated (CAND1) modulates SCF function through its interactions with the CUL1 subunit. Although biochemical studies with human CAND1 suggested that CAND1 plays a negative regulatory role by sequestering CUL1 and preventing SCF complex assembly, genetic studies in Arabidopsis have shown that cand1 mutants exhibit reduced SCF activity, demonstrating that CAND1 is required for optimal SCF function in vivo. Together, these genetic and biochemical studies have suggested a model of CAND1-mediated cycles of SCF complex assembly and disassembly. Here, using the SCF(TIR1) complex of the Arabidopsis auxin response pathway, we test the SCF cycling model with Arabidopsis mutant derivatives of CAND1 and CUL1 that have opposing effects on the CAND1-CUL1 interaction. We find that the disruption of the CAND1-CUL1 interaction results in an increased abundance of assembled SCF(TIR1) complex. In contrast, stabilization of the CAND1-CUL1 interaction diminishes SCF(TIR1) complex abundance. The fact that both decreased and increased CAND1-CUL1 interactions result in reduced SCF(TIR1) activity in vivo strongly supports the hypothesis that CAND1-mediated cycling is required for optimal SCF function.
Zhang
Wenjing
W
Department of Plant Biology, University of Minnesota-Twin Cities, St. Paul, MN 55108.
Ito
Hironori
H
Quint
Marcel
M
Huang
He
H
Noël
Laurent D
LD
Gray
William M
WM
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D044842:Cullin Proteins / Q000235:genetics / Q000378:metabolism*; D004252:DNA Mutational Analysis; D009154:Mutation; D044843:SKP Cullin F-Box Protein Ligases / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D029681:Arabidopsis Proteins; C488785:CAND1 protein, Arabidopsis; C100318:Cullin 1; D044842:Cullin Proteins; D044843:SKP Cullin F-Box Protein Ligases