Two steps of insulin receptor internalization depend on different domains of the beta-subunit.
The internalization of signaling receptors such as the insulin receptor is a complex, multi-step process. The aim of the present work was to determine the various steps in internalization of the insulin receptor and to establish which receptor domains are implicated in each of these by the use of receptors possessing in vitro mutations. We find that kinase activation and autophosphorylation of all three regulatory tyrosines 1146, 1150, and 1151, but not tyrosines 1316 and 1322 in the COOH-terminal domain, are required for the ligand-specific stage of the internalization process; i.e., the surface redistribution of the receptor from microvilli where initial binding occurs to the nonvillous domain of the cell. Early intracellular steps in insulin signal transduction involving the activation of phosphatidylinositol 3'-kinase are not required for this redistribution. The second step of internalization consists in the anchoring of the receptors in clathrin-coated pits. In contrast to the first ligand specific step, this step is common to many receptors including those for transport proteins and occurs in the absence of kinase activation and receptor autophosphorylation, but requires a juxta-membrane cytoplasmic segment of the beta-subunit of the receptor including a NPXY sequence. Thus, there are two independent mechanisms controlling insulin receptor internalization which depend on different domains of the beta-subunit.
Department of Morphology, University of Geneva, Switzerland.
J L
JL
Carpentier
J P
JP
Paccaud
J
J
Backer
A
A
Gilbert
L
L
Orci
C R
CR
Kahn
J ]
J]
Baecker J [corrected to Backer
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D016466:CHO Cells; D002966:Clathrin / Q000378:metabolism; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism / Q000648:ultrastructure; D006224:Cricetinae; D007328:Insulin / Q000032:analysis / Q000378:metabolism; D007457:Iodine Radioisotopes; D008871:Microvilli / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D011972:Receptor, Insulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
D002966:Clathrin; D007328:Insulin; D007457:Iodine Radioisotopes; D011972:Receptor, Insulin