EHD2 and the novel EH domain binding protein EHBP1 couple endocytosis to the actin cytoskeleton.
Here we identified two novel proteins denoted EH domain protein 2 (EHD2) and EHD2-binding protein 1 (EHBP1) that link clathrin-mediated endocytosis to the actin cytoskeleton. EHD2 contains an N-terminal P-loop and a C-terminal EH domain that interacts with NPF repeats in EHBP1. Disruption of EHD2 or EHBP1 function by small interfering RNA-mediated gene silencing inhibits endocytosis of transferrin into EEA1-positive endosomes as well as GLUT4 endocytosis into cultured adipocytes. EHD2 localizes with cortical actin filaments, whereas EHBP1 contains a putative actin-binding calponin homology domain. High expression of EHD2 or EHBP1 in intact cells mediates extensive actin reorganization. Thus EHD2 appears to connect endocytosis to the actin cytoskeleton through interactions of its N-terminal domain with membranes and its C-terminal EH domain with the novel EHBP1 protein.
Guilherme
Adilson
A
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Soriano
Neil A
NA
Bose
Sahana
S
Holik
John
J
Bose
Avirup
A
Pomerleau
Darcy P
DP
Furcinitti
Paul
P
Leszyk
John
J
Corvera
Silvia
S
Czech
Michael P
MP
D041721:3T3-L1 Cells; D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D017667:Adipocytes / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D015152:Blotting, Northern; D015153:Blotting, Western; D019556:COS Cells; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D020868:Gene Silencing; D005947:Glucose / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008957:Models, Genetic; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D012235:Rhodamines / Q000737:chemistry; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D014168:Transferrin / Q000737:chemistry / Q000378:metabolism
D000199:Actins; D002352:Carrier Proteins; D018076:DNA, Complementary; C485118:EHBP1 protein, human; C485117:EHD2 protein, mouse; C484942:EHD2-binding protein 1, mouse; D008164:Luminescent Proteins; D034741:RNA, Small Interfering; D012235:Rhodamines; D014168:Transferrin; D049452:Green Fluorescent Proteins; D005947:Glucose