Actin dynamics coupled to clathrin-coated vesicle formation at the trans-Golgi network.
In diverse species, actin assembly facilitates clathrin-coated vesicle (CCV) formation during endocytosis. This role might be an adaptation specific to the unique environment at the cell cortex, or it might be fundamental, facilitating CCV formation on different membranes. Proteins of the Sla2p/Hip1R family bind to actin and clathrin at endocytic sites in yeast and mammals. We hypothesized that Hip1R might also coordinate actin assembly with clathrin budding at the trans-Golgi network (TGN). Using deconvolution and time-lapse microscopy, we showed that Hip1R is present on CCVs emerging from the TGN. These vesicles contain the mannose 6-phosphate receptor involved in targeting proteins to the lysosome, and the actin nucleating Arp2/3 complex. Silencing of Hip1R expression by RNAi resulted in disruption of Golgi organization and accumulation of F-actin structures associated with CCVs on the TGN. Hip1R silencing and actin poisons slowed cathepsin D exit from the TGN. These studies establish roles for Hip1R and actin in CCV budding from the TGN for lysosome biogenesis.
Carreno
Sebastien
S
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3202, USA.
Engqvist-Goldstein
Asa E
AE
Zhang
Claire X
CX
McDonald
Kent L
KL
Drubin
David G
DG
D000199:Actins / Q000502:physiology*; D048868:Adaptor Proteins, Signal Transducing; D002402:Cathepsin D / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000502:physiology* / Q000648:ultrastructure; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008247:Lysosomes / Q000502:physiology; D011485:Protein Binding; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D021601:trans-Golgi Network / Q000502:physiology* / Q000648:ultrastructure
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D034741:RNA, Small Interfering; C441328:SLA2 protein, human; D016392:Proto-Oncogene Proteins pp60(c-src); D002402:Cathepsin D