Activation of insulin receptor signaling by a single amino acid substitution in the transmembrane domain.
The insulin receptor is a ligand-activated tyrosine kinase composed of two alpha and two beta subunits. A single transmembrane domain composed of 23 hydrophobic residues is contained in each beta subunit. We examined the role of the transmembrane domain in regulating insulin receptor signaling by inserting a negatively charged amino acid (Asp) for Val938 (V938D). Chinese hamster ovary (CHO) cells were stably transfected with a plasmid containing both the neomycin-resistance gene and either the wild-type or the mutant (V938D) insulin receptor cDNA. Insulin binding increased similarly in CHO cells stably transfected with the wild-type and the V938D-mutant insulin receptor cDNA. Insulin stimulated glucose transport and cell growth in cells expressing the normal insulin receptor. By contrast, in the absence of insulin, glucose transport and cell growth in CHO-V938D cells were as high as in insulin-stimulated control cells and no longer responsive to insulin stimulation. Phosphorylation of the beta subunit of the insulin receptor was also increased in CHO-V938D cells not exposed to insulin. These results support an essential role of the transmembrane domain of the insulin receptor in the transduction of insulin signaling.
Longo
N
N
Department of Pediatrics, Emory University, Atlanta, Georgia 30322.
Shuster
R C
RC
Griffin
L D
LD
Langley
S D
SD
Elsas
L J
LJ
D000818:Animals; D001216:Asparagine / Q000378:metabolism; D001483:Base Sequence; D001692:Biological Transport; D016466:CHO Cells; D002455:Cell Division; D006224:Cricetinae; D004247:DNA; D004789:Enzyme Activation; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011972:Receptor, Insulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism; D014633:Valine / Q000378:metabolism
D007328:Insulin; D014443:Tyrosine; D001216:Asparagine; D004247:DNA; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D014633:Valine; D005947:Glucose