Present address: Agriculture and Agri-Food Canada, Cereal Research Centre, 195 Dafoe Road, Winnipeg, MB, Canada R3T-2M9.
Protein interaction analysis of SCF ubiquitin E3 ligase subunits from Arabidopsis
Article first published online: 10 JUN 2003
The Plant Journal
Volume 34, Issue 6, pages 753–767, June 2003
How to Cite
Risseeuw, E. P., Daskalchuk, T. E., Banks, T. W., Liu, E., Cotelesage, J., Hellmann, H., Estelle, M., Somers, D. E. and Crosby, W. L. (2003), Protein interaction analysis of SCF ubiquitin E3 ligase subunits from Arabidopsis. The Plant Journal, 34: 753–767. doi: 10.1046/j.1365-313X.2003.01768.x
- Issue published online: 10 JUN 2003
- Article first published online: 10 JUN 2003
- Received 20 September 2002; revised 28 January 2003; accepted 5 February 2003.
- E3 ligase;
- F-box gene family;
- yeast two-hybrid;
- ASK gene family
Ubiquitin E3 ligases are a diverse family of protein complexes that mediate the ubiquitination and subsequent proteolytic turnover of proteins in a highly specific manner. Among the several classes of ubiquitin E3 ligases, the Skp1-Cullin-F-box (SCF) class is generally comprised of three ‘core’ subunits: Skp1 and Cullin, plus at least one F-box protein (FBP) subunit that imparts specificity for the ubiquitination of selected target proteins. Recent genetic and biochemical evidence in Arabidopsis thaliana suggests that post-translational turnover of proteins mediated by SCF complexes is important for the regulation of diverse developmental and environmental response pathways. In this report, we extend upon a previous annotation of the Arabidopsis Skp1-like (ASK) and FBP gene families to include the Cullin family of proteins. Analysis of the protein interaction profiles involving the products of all three gene families suggests a functional distinction between ASK proteins in that selected members of the protein family interact generally while others interact more specifically with members of the F-box protein family. Analysis of the interaction of Cullins with FBPs indicates that CUL1 and CUL2, but not CUL3A, persist as components of selected SCF complexes, suggesting some degree of functional specialization for these proteins. Yeast two-hybrid analyses also revealed binary protein interactions between selected members of the FBP family in Arabidopsis. These and related results are discussed in terms of their implications for subunit composition, stoichiometry and functional diversity of SCF complexes in Arabidopsis.