Abstract
Regulated proteolysis by the ubiquitin-26S proteasome system challenges transcription and phosphorylation in magnitude and is one of the most important regulatory mechanisms in plants. This article describes the characterization of a rice (Oryza sativa) auxin-responsive Kelch-domain-containing F-box protein, OsFBK1, found to be a component of an SCF E3 ligase by interaction studies in yeast. Rice transgenics of OsFBK1 displayed variations in anther and root secondary cell wall content; it could be corroborated by electron/confocal microscopy and lignification studies, with no apparent changes in auxin content/signaling pathway. The presence of U-shaped secondary wall thickenings (or lignin) in the anthers were remarkably less pronounced in plants overexpressing OsFBK1 as compared to wild-type and knockdown transgenics. The roots of the transgenics also displayed differential accumulation of lignin. Yeast two-hybrid anther library screening identified an OsCCR that is a homolog of the well-studied Arabidopsis (Arabidopsis thaliana) IRX4; OsFBK1-OsCCR interaction was confirmed by fluorescence and immunoprecipitation studies. Degradation of OsCCR mediated by SCFOsFBK1 and the 26S proteasome pathway was validated by cell-free experiments in the absence of auxin, indicating that the phenotype observed is due to the direct interaction between OsFBK1 and OsCCR. Interestingly, the OsCCR knockdown transgenics also displayed a decrease in root and anther lignin depositions, suggesting that OsFBK1 plays a role in the development of rice anthers and roots by regulating the cellular levels of a key enzyme controlling lignification.