TY - JOUR
T1 - Phosphatidylinositol-4,5-bisphosphate is required for endocytic coated vesicle formation
AU - Jost, Matthias
AU - Simpson, Fiona
AU - Kavran, Jennifer M.
AU - Lemmon, Mark A.
AU - Schmid, Sandra L.
N1 - Funding Information:
We thank M. Fujimoto for technical help with the BSSTfn (MesNa) assay, and C. Alory, H. Damke and K. Fish for comments on the manuscript. M.J. was supported by DFG fellowship Jo282/1-1. F.S. was supported by Well-come Trust Fellowship 049043/Z96/ZJMW/LEC. M.A.L. was supported by NIGMS grant GM56846 and by a Scholar Award (DRS-05) from the Damon Runyon-Walter Winchell Foundation. S.L.S. was supported by NCI grant CA69099 and is an Established Investigator of the American Heart Association. This is TSRI manuscript No. 11964-CB.
PY - 1998/12/31
Y1 - 1998/12/31
N2 - Receptor-mediated endocytosis via clathrin-coated vesicles has been extensively studied and, while many of the protein players have been identified, much remains unknown about the regulation of coat assembly and the mechanisms that drive vesicle formation [1]. Some components of the endocytic machinery interact with inositol polyphosphates and inositol lipids in vitro, implying a role for phosphatidylinositols in vivo [2,3]. Specifically, the adaptor protein complex AP2 binds phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2), PtdIns(3)P, PtdIns(3,4,5)P3 and inositol phosphates. Phosphatidylinositol binding regulates AP2 self-assembly and the interactions of AP2 complexes with clathrin and with peptides containing endocytic motifs [4,5]. The GTPase dynamin contains a pleckstrin homology (PH) domain that binds PtdIns(4,5)P2 and PtdIns(3,4,5)P3 to regulate GTPase activity in vitro [6,7]. However, no direct evidence for the involvement of phosphatidylinositols in clathrin-mediated endocytosis exists to date. Using well-characterized PH domains as high affinity and high specificity probes in combination with a perforated cell assay that reconstitutes coated vesicle formation, we provide the first direct evidence that PtdIns(4,5)P2 is required for both early and late events in endocytic coated vesicle formation.
AB - Receptor-mediated endocytosis via clathrin-coated vesicles has been extensively studied and, while many of the protein players have been identified, much remains unknown about the regulation of coat assembly and the mechanisms that drive vesicle formation [1]. Some components of the endocytic machinery interact with inositol polyphosphates and inositol lipids in vitro, implying a role for phosphatidylinositols in vivo [2,3]. Specifically, the adaptor protein complex AP2 binds phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2), PtdIns(3)P, PtdIns(3,4,5)P3 and inositol phosphates. Phosphatidylinositol binding regulates AP2 self-assembly and the interactions of AP2 complexes with clathrin and with peptides containing endocytic motifs [4,5]. The GTPase dynamin contains a pleckstrin homology (PH) domain that binds PtdIns(4,5)P2 and PtdIns(3,4,5)P3 to regulate GTPase activity in vitro [6,7]. However, no direct evidence for the involvement of phosphatidylinositols in clathrin-mediated endocytosis exists to date. Using well-characterized PH domains as high affinity and high specificity probes in combination with a perforated cell assay that reconstitutes coated vesicle formation, we provide the first direct evidence that PtdIns(4,5)P2 is required for both early and late events in endocytic coated vesicle formation.
UR - http://www.scopus.com/inward/record.url?scp=0032585530&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032585530&partnerID=8YFLogxK
U2 - 10.1016/s0960-9822(98)00022-0
DO - 10.1016/s0960-9822(98)00022-0
M3 - Article
C2 - 9889104
AN - SCOPUS:0032585530
SN - 0960-9822
VL - 8
SP - 1399
EP - 1404
JO - Current Biology
JF - Current Biology
IS - 25
ER -