Genetic analysis of endocytosis in Caenorhabditis elegans: coelomocyte uptake defective mutants.
The coelomocytes of Caenorhabditis elegans are scavenger cells that continuously and nonspecifically endocytose fluid from the pseudocoelom (body cavity). Green fluorescent protein (GFP) secreted into the pseudocoelom from body wall muscle cells is endocytosed and degraded by coelomocytes. We show that toxin-mediated ablation of coelomocytes results in viable animals that fail to endocytose pseudocoelomic GFP, indicating that endocytosis by coelomocytes is not essential for growth or survival of C. elegans under normal laboratory conditions. We examined known viable endocytosis mutants, and performed RNAi for other known endocytosis genes, for coelomocyte uptake defective (Cup) phenotypes. We also screened for new genes involved in endocytosis by isolating viable mutants with Cup defects; this screen identified 14 different genes, many with multiple alleles. A variety of Cup terminal phenotypes were observed, consistent with defects at various steps in the endocytic pathway. Available molecular information indicates that the Cup mutant screen has identified novel components of the endocytosis machinery that are conserved in mammals but not in Saccharomyces cerevisiae, the only other organism for which large-scale genetic screens for endocytosis mutants have been performed.
Fares
H
H
Department of Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA. fares@email.uarizona.edu
Greenwald
I
I
D000483:Alleles; D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000235:genetics* / Q000502:physiology*; D004705:Endocytosis / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006358:Hot Temperature; D008164:Luminescent Proteins / Q000378:metabolism / Q000493:pharmacokinetics; D008953:Models, Anatomic; D009154:Mutation; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012329:RNA, Bacterial / Q000378:metabolism; D011995:Recombination, Genetic; D013997:Time Factors; D019076:Transgenes
D008164:Luminescent Proteins; C042027:RNA I; D012329:RNA, Bacterial; D049452:Green Fluorescent Proteins; D012313:RNA