unc-41stonin functions with ap2 to recycle synaptic vesicles in caenorhabditis elegansunc-41stonin函数ap2回收突触囊泡在秀丽隐杆线虫.pdfVIP

UNC-41/Stonin Functions with AP2 to Recycle Synaptic Vesicles in Caenorhabditis elegans 1. 1. 2. 2 2 Gregory P. Mullen , Kiely M. Grundahl , Mingyu Gu , Shigeki Watanabe , Robert J. Hobson , 1{ 1 1 2 1 John A. Crowell , John R. McManus , Eleanor A. Mathews , Erik M. Jorgensen , James B. Rand * 1 Genetic Models of Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, United States of America, 2 Howard Hughes Medical Institute and Department of Biology, University of Utah, Salt Lake City, Utah, United States of America Abstract The recycling of synaptic vesicles requires the recovery of vesicle proteins and membrane. Members of the stonin protein family (Drosophila Stoned B, mammalian stonin 2) have been shown to link the synaptic vesicle protein synaptotagmin to the endocytic machinery. Here we characterize the unc-41 gene, which encodes the stonin ortholog in the nematode Caenorhabditis elegans. Transgenic expression of Drosophila stonedB rescues unc-41 mutant phenotypes, demonstrating that UNC-41 is a bona fide member of the stonin family. In unc-41 mutants, synaptotagmin is present in axons, but is mislocalized and diffuse. In contrast, UNC-41 is localized normally in synaptotagmin mutants, demonstrating a unidirectional relationship for localization. The phenotype of snt-1 unc-41 double mutants is stronger than snt-1 mutants, suggesting that UNC-41 may have additional, synaptotagmin-independent functions. We also show that unc-41 mutants have defects in synaptic vesicle membrane endocytosis, including a ,50%