EEA1 links PI(3)K function to Rab5 regulation of endosome fusion.
GTPases and lipid kinases regulate membrane traffic along the endocytic pathway by mechanisms that are not completely understood. Fusion between early endosomes requires phosphatidylinositol-3-OH kinase (PI(3)K) activity as well as the small GTPase Rab5. Excess Rab5-GTP complex restores endosome fusion when PI(3)K is inhibited. Here we identify the early-endosomal autoantigen EEA1 which binds the PI(3)K product phosphatidylinositol-3-phosphate, as a new Rab5 effector that is required for endosome fusion. The association of EEA1 with the endosomal membrane requires Rab5-GTP and PI(3)K activity, and excess Rab5-GTP stabilizes the membrane association of EEA1 even when PI(3)K is inhibited. The identification of EEA1 as a direct Rab5 effector provides a molecular link between PI(3)K and Rab5, and its restricted distribution to early endosomes indicates that EEA1 may confer directionality to Rab5-dependent endocytic transport.
Department of Biochemistry, The Norwegian Radium Hospital, Montebello, Oslo.
A
A
Simonsen
R
R
Lippé
S
S
Christoforidis
J M
JM
Gaullier
A
A
Brech
J
J
Callaghan
B H
BH
Toh
C
C
Murphy
M
M
Zerial
H
H
Stenmark
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001324:Autoantigens / Q000502:physiology; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D011992:Endosomes / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*; D006160:Guanosine Triphosphate / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000502:physiology; D008561:Membrane Fusion / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D016296:Mutagenesis; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D033921:Vesicular Transport Proteins; D000077191:Wortmannin; D020696:rab5 GTP-Binding Proteins
D000730:Androstadienes; D001324:Autoantigens; D004791:Enzyme Inhibitors; D019204:GTP-Binding Proteins; D006160:Guanosine Triphosphate; D008565:Membrane Proteins; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D033921:Vesicular Transport Proteins; D000077191:Wortmannin; D020696:rab5 GTP-Binding Proteins; C093846:early endosome antigen 1