Yeast Eps15-like endocytic protein, Pan1p, activates the Arp2/3 complex.
Longstanding evidence supports a role for actin in endocytosis; an intact actin cytoskeleton is required for endocytosis in yeast, and drugs that inhibit actin polymerization inhibit endocytosis in both yeast and mammalian cells. The yeast Arp2/3 complex is required for the internalization step of endocytosis. In addition, some early endocytic events in mammalian cells are associated with the formation of actin tails similar to those generated by activated Arp2/3 complex. However, until now no Arp2/3 complex activator has been identified among proteins known to mediate early steps in endocytosis. Here we show that the yeast endocytic protein Pan1p binds to and activates the Arp2/3 complex. Genetic interactions between PAN1 and mutants of Arp2/3 subunits, or of the Arp2/3 activator LAS17, provide evidence for this activity in vivo. We suggest that Pan1p forms the core of an endocytic complex and physically couples actin polymerization nucleated by the Arp2/3 complex to the endocytic machinery, thus providing the forces necessary for endocytosis.
Duncan
M C
MC
Department of Molecular and Cell Biology, 401 Barker Hall, University of California, Berkeley, California 94720-3202, USA.
Cope
M J
MJ
Goode
B L
BL
Wendland
B
B
Drubin
D G
DG
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000187:drug effects* / Q000378:metabolism; D000595:Amino Acid Sequence; D002135:Calcium-Binding Proteins / Q000502:physiology*; D003598:Cytoskeletal Proteins; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000187:drug effects; D005656:Fungal Proteins / Q000235:genetics / Q000494:pharmacology*; D007700:Kinetics; D008840:Microfilament Proteins; D010750:Phosphoproteins / Q000502:physiology*; D011485:Protein Binding; D012440:Saccharomyces / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017384:Sequence Deletion
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; D002135:Calcium-Binding Proteins; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; D008840:Microfilament Proteins; C111870:PAN1 protein, S cerevisiae; D010750:Phosphoproteins; D029701:Saccharomyces cerevisiae Proteins