The phosphorylation of an actin depolymerizing factor by a calcium-dependent protein kinase regulates cotton fiber elongation

A cotton fiber is a single and highly elongated ovule epidermal cell. However, the mechanism that governs the development of fiber traits remains unclear. In this study, we cloned a calcium-dependent protein kinase (GhCPK1) and an actin depolymerizing factor (GhADF1) from Gossypium hirsutum. Real-time PCR analyses indicated that the expression of GhCPK1 and GhADF1 correlated with the elongation pattern of cotton fibers. Yeast two-hybrid assays using full-length GhCPK1 and truncated forms of GhCPK1 as baits identified GhADF1 as an interactor of GhCPK1. Furthermore, GhCPK1 is capable of phosphorylating GhADF1 in vitro in a calcium-dependent manner, and the phosphorylation of GhADF1 by GhCPK1 occurs on the Ser-6 of GhADF1. In addition, we observed that the heterologous expression of the GhCPK1 gene induced longitudinal growth of the host cells by 3.18-fold, with no apparent effect on other aspects of the host cells. The results strongly suggest that GhCPK1 may regulate the function of GhADF1 by phosphorylating this protein during cotton fiber elongation.