Independent roles of Arp2/3 complex and RIC4 protein in the control of epidermal cell shape

The aim of this study was to determine whether RIC4, a CRIB domain-containing protein that functions as an effector of ROP GTPases, and the Arp2/3 complex, an actin nucleator, functionally cooperate in controlling the shape of Arabidopsis cotyledon epidermal cells. The combination of KO mutants demonstrated that loss of RIC4 and loss of the Arp2/3 complex result in completely opposite epidermal cell shape phenotypes. The double KO mutation is phenotypically similar to the Arp2/3 mutation, and the effect of RIC4 loss is completely eliminated. Analysis of overexpression revealed that excess RIC4 significantly suppresses the formation of pavement cell lobes. However, RIC4 does not require an active Arp2/3 complex for this effect. Our data further show that overexpression of RIC4 has a specific actin stabilization effect in cotyledon epidermal cells. However, we could not demonstrate that actin stabilization is directly related to the cell shape changes that RIC4 overexpression induces. In conclusion, RIC4 and the Arp2/3 complex do not share the same signaling pathway in their function in the control of cotyledon epidermal cell shape.