Control of clathrin-mediated endocytosis by NIMA family kinases

Autoři: Braveen B. Joseph aff001;  Yu Wang aff002;  Phil Edeen aff001;  Vladimir Lažetić aff001;  Barth D. Grant aff002;  David S. Fay aff001
Působiště autorů: Department of Molecular Biology, College of Agriculture and Natural Resources, University of Wyoming, Laramie, Wyoming, United States of America aff001;  Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey, United States of America aff002;  Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China aff003
Vyšlo v časopise: Control of clathrin-mediated endocytosis by NIMA family kinases. PLoS Genet 16(2): e32767. doi:10.1371/journal.pgen.1008633
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008633


Endocytosis, the process by which cells internalize plasma membrane and associated cargo, is regulated extensively by posttranslational modifications. Previous studies suggested the potential involvement of scores of protein kinases in endocytic control, of which only a few have been validated in vivo. Here we show that the conserved NIMA-related kinases NEKL-2/NEK8/9 and NEKL-3/NEK6/7 (the NEKLs) control clathrin-mediated endocytosis in C. elegans. Loss of NEKL-2 or NEKL-3 activities leads to penetrant larval molting defects and to the abnormal localization of trafficking markers in arrested larvae. Using an auxin-based degron system, we also find that depletion of NEKLs in adult-stage C. elegans leads to gross clathrin mislocalization and to a dramatic reduction in clathrin mobility at the apical membrane. Using a non-biased genetic screen to identify suppressors of nekl molting defects, we identified several components and regulators of AP2, the major clathrin adapter complex acting at the plasma membrane. Strikingly, reduced AP2 activity rescues both nekl mutant molting defects as well as associated trafficking phenotypes, whereas increased levels of active AP2 exacerbate nekl defects. Moreover, in a unique example of mutual suppression, NEKL inhibition alleviates defects associated with reduced AP2 activity, attesting to the tight link between NEKL and AP2 functions. We also show that NEKLs are required for the clustering and internalization of membrane cargo required for molting. Notably, we find that human NEKs can rescue molting and trafficking defects in nekl mutant worms, suggesting that the control of intracellular trafficking is an evolutionarily conserved function of NEK family kinases.

Klíčová slova:

Auxins – Caenorhabditis elegans – Cell membranes – Endocytosis – Fluorescence recovery after photobleaching – Larvae – Molting – RNA interference


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