Interactions between Piccolo and the actin/dynamin-binding protein Abp1 link vesicle endocytosis to presynaptic active zones.
Piccolo is a high molecular weight multi-domain protein shown to be a structural component of the presynaptic CAZ (cytoskeletal matrix assembled at active zones). These features indicate that Piccolo may act to scaffold proteins involved in synaptic vesicle endo- and exocytosis near their site of action. To test this hypothesis, we have utilized a functional cell-based endocytosis assay and identified the N-terminal proline-rich Q domain in Piccolo as a region that interferes with clathrin-mediated endocytosis. Utilizing the Piccolo Q domain as bait in a yeast two-hybrid screen, we have identified the F-actin-binding protein Abp1 (also called SH3P7 or HIP-55) as a potential binding partner for this domain. The physiological relevance of this interaction is supported by in vitro binding studies, colocalization in nerve terminals, in vivo recruitment studies, and immunoprecipitation experiments. Intriguingly, Abp1 binds to both F-actin and the GTPase dynamin and has been implicated in linking the actin cytoskeleton to clathrin-mediated endocytosis. Our results suggest that Piccolo, as a structural protein of the CAZ, may serve to localize Abp1 at active zones where it can actively participate in creating a functional connection between the dynamic actin cytoskeleton and synaptic vesicle recycling.
Fenster
Steven D
SD
Department of Neurobiology, Civitan International Research Center, University of Alabama at Birmingham, 35294, USA.
Kessels
Michael M
MM
Qualmann
Britta
B
Chung
Wook J
WJ
Nash
Joanne
J
Gundelfinger
Eckart D
ED
Garner
Craig C
CC
D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004705:Endocytosis; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism*; D011233:Precipitin Tests; D017729:Presynaptic Terminals / Q000378:metabolism*; D011485:Protein Binding; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
C062612:ABP1 protein, S cerevisiae; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D009479:Neuropeptides; C405514:Pclo protein, rat; D029701:Saccharomyces cerevisiae Proteins