Neural Wiskott Aldrich Syndrome Protein (N-WASP) and the Arp2/3 complex are recruited to sites of clathrin-mediated endocytosis in cultured fibroblasts

CJ Merrifield, B Qualmann, MM Kessels… - European journal of cell …, 2004 - Elsevier
CJ Merrifield, B Qualmann, MM Kessels, W Almers
European journal of cell biology, 2004Elsevier
Several findings suggest that actin-mediated motility can play a role in clathrin-mediated
endocytosis but it remains unclear whether and when key proteins required for this process
are recruited to endocytic sites. Here we investigate this question in live Swiss 3T3 cells
using two-colour evanescent field (EF) microscopy. We find that Arp3, a component of the
Arp2/3 complex, appears transiently while single clathrin-coated pits internalize. There is
also additional recruitment of Neural-Wiskott Aldrich Syndrome Protein (N-WASP), a known …
Summary
Several findings suggest that actin-mediated motility can play a role in clathrin-mediated endocytosis but it remains unclear whether and when key proteins required for this process are recruited to endocytic sites. Here we investigate this question in live Swiss 3T3 cells using two-colour evanescent field (EF) microscopy. We find that Arp3, a component of the Arp2/3 complex, appears transiently while single clathrin-coated pits internalize. There is also additional recruitment of Neural-Wiskott Aldrich Syndrome Protein (N-WASP), a known activator of the Arp2/3 complex. Both proteins appear at about the same time as actin. We suggest that N-WASP and the Arp2/3 complex trigger actin polymerization during a late step in clathrin-mediated endocytosis, and propel clathrin-coated pits or vesicles from the plasma membrane into the cytoplasm.
Elsevier