Journal
NATURE CELL BIOLOGY
Volume 14, Issue 7, Pages 666-676Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/ncb2508
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Funding
- German Federal Ministry of Education and Research (InnoRegio Initiative) [03I4035A]
- German Federal Ministry of Education and Research (Systems Biology Network HepatoSys) [0313082H]
- German Federal Ministry of Education and Research (Virtual Liver initiative)
- EU [LSHG-CT-2006-019050]
- Max Planck Society's inter-institutional initiative 'RNA interference'
- Boehringer Ingelheim
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Apically enriched Rab11-positive recycling endosomes (Rab11-REs) are important for establishing and maintaining epithelial polarity. Yet, little is known about the molecules controlling trafficking of Rab11-REs in an epithelium in vivo. Here, we report a genome-wide, image-based RNA interference screen for regulators of Rab11-RE positioning and transport of an apical membrane protein (PEPT-1) in C. elegans intestine. Among the 356 screen hits was the 14-3-3 and partitioning defective protein PAR-5, which we found to be specifically required for Rab11-RE positioning and apicobasal polarity maintenance. Depletion of PAR-5 induced abnormal clustering of Rab11-REs to ectopic sites at the basolateral cortex containing F-actin and other apical domain components. This phenotype required key regulators of F-actin dynamics and polarity, such as Rho GTPases (RHO-1 and the Rac1 orthologue CED-10) and apical PAR proteins. Our data suggest that PAR-5 acts as a regulatory hub for a polarity-maintaining network required for apicobasal asymmetry of F-actin and proper Rab11-RE positioning.
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