Control of clathrin-mediated endocytosis by NIMA family kinases

PLoS Genet. 2020 Feb 18;16(2):e1008633. doi: 10.1371/journal.pgen.1008633. eCollection 2020 Feb.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Adaptor Protein Complex 2 / metabolism
  • Animals
  • Animals, Genetically Modified
  • Caenorhabditis elegans / physiology*
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism
  • Cell Membrane / metabolism
  • Clathrin / metabolism*
  • Endocytosis
  • Intravital Microscopy
  • Larva / growth & development
  • Molting / genetics
  • Mutation
  • NIMA-Related Kinases / genetics*
  • NIMA-Related Kinases / metabolism
  • Protein Kinases / genetics*
  • Protein Kinases / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Time-Lapse Imaging


  • Adaptor Protein Complex 2
  • Caenorhabditis elegans Proteins
  • Clathrin
  • NEKL-3 protein, C elegans
  • Recombinant Proteins
  • Protein Kinases
  • NEK6 protein, human
  • NEK7 protein, human
  • NEKL-2 protein, C elegans
  • NIMA-Related Kinases