Journal
EUROPEAN JOURNAL OF CELL BIOLOGY
Volume 83, Issue 1, Pages 13-18Publisher
ELSEVIER GMBH, URBAN & FISCHER VERLAG
DOI: 10.1078/0171-9335-00356
Keywords
actin mediated motility; two-colour evanescent field microscopy; protein interactions in vivo
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Funding
- NIDDK NIH HHS [P01 DK44239] Funding Source: Medline
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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 rind 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 a ut 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.
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