The Disease-Associated Formin INF2/EXC-6 Organizes Lumen and Cell Outgrowth during Tubulogenesis by Regulating F-Actin and Microtubule Cytoskeletons

• C. elegans EXC-6 is orthologous to the vertebrate formin INF2
• INF2 disease-causing mutations are constitutively active in tubulogenesis in worms
• EXC-6/INF2 regulates F-actin accumulation at the tip of growing tubes
• EXC-6/INF2 regulates the dynamics of basolateral-localized MTs in growing tubes

SummaryWe investigate how outgrowth at the basolateral cell membrane is coordinated with apical lumen formation in the development of a biological tube by characterizing exc-6, a gene required for C. elegans excretory cell (EC) tubulogenesis. We show that EXC-6 is orthologous to the human formin INF2, which polymerizes filamentous actin (F-actin) and binds microtubules (MTs) in vitro. Dominant INF2 mutations cause focal segmental glomerulosclerosis (FSGS), a kidney disease, and FSGS+Charcot-Marie-Tooth neuropathy. We show that activated INF2 can substitute for EXC-6 in C. elegans and that disease-associated mutations cause constitutive activity. Using genetic analysis and live imaging, we show that exc-6 regulates MT and F-actin accumulation at EC tips and dynamics of basolateral-localized MTs, indicating that EXC-6 organizes F-actin and MT cytoskeletons during tubulogenesis. The pathology associated with INF2 mutations is believed to reflect misregulation of F-actin, but our results suggest alternative or additional mechanisms via effects on MT dynamics.