Requirements for F-BAR Proteins TOCA-1 and TOCA-2 in Actin Dynamics and Membrane Trafficking during Caenorhabditis elegans Oocyte Growth and Embryonic Epidermal Morphogenesis

Abstract

The TOCA family of F-BAR–containing proteins bind to and remodel lipid bilayers via their conserved F-BAR domains, and regulate actin dynamics via their N-Wasp binding SH3 domains. Thus, these proteins are predicted to play a pivotal role in coordinating membrane traffic with actin dynamics during cell migration and tissue morphogenesis. By combining genetic analysis in Caenorhabditis elegans with cellular biochemical experiments in mammalian cells, we showed that: i) loss of CeTOCA proteins reduced the efficiency of Clathrin-mediated endocytosis (CME) in oocytes. Genetic interference with CeTOCAs interacting proteins WSP-1 and WVE-1, and other components of the WVE-1 complex, produced a similar effect. Oocyte endocytosis defects correlated well with reduced egg production in these mutants. ii) CeTOCA proteins localize to cell–cell junctions and are required for proper embryonic morphogenesis, to position hypodermal cells and to organize junctional actin and the junction-associated protein AJM-1. iii) Double mutant analysis indicated that the toca genes act in the same pathway as the nematode homologue of N-WASP/WASP, wsp-1. Furthermore, mammalian TOCA-1 and C. elegans CeTOCAs physically associated with N-WASP and WSP-1 directly, or WAVE2 indirectly via ABI-1. Thus, we propose that TOCA proteins control tissues morphogenesis by coordinating Clathrin-dependent membrane trafficking with WAVE and N-WASP–dependent actin-dynamics.

Author Summary

Cells continuously remodel their shape especially during cell migration, differentiation, and tissues morphogenesis. This occurs through the dynamic reorganization of their plasma membrane and actin cytoskeleton: two processes that must therefore be intimately linked and coordinated. Molecules that sit at the crossroads of membrane remodeling and actin dynamics are predicted to play a pivotal role in coordinating these processes. The TOCA family of proteins represents a case in point. These proteins bind to and deform membranes during processes such as membrane trafficking. They also control actin dynamics through their interactions with actin remodeling factors, such as WASP and WAVEs. Here, we characterize the functional role of TOCA proteins in a model organism, the nematode Caenorhabditis elegans. We established that toca genes regulate Clathrin-mediated membrane trafficking during oocyte growth. We further discovered that these proteins play an important role in epithelial morphogenesis in developing embryos, and in egg production in adult nematodes. Moreover, the TOCA interacting proteins WASP/WSP-1 and WAVE/WVE-1, as well as other components of the WVE-1 complex, appear to be involved in TOCA-dependent processes. Thus, we propose that TOCA proteins control tissue morphogenesis by coordinating Clathrin-dependent membrane trafficking with WAVE and N-WASP–dependent actin-dynamics.