The adapter protein GRB10 is an endogenous negative regulator of insulin-like growth factor signaling.
The growth factor IGF-I is critical for normal human somatic growth and development. Growth factor receptor-bound protein (Grb)10 is a protein that interacts with the IGF-I receptor and may thus regulate IGF-I-stimulated growth. However, the role of endogenous Grb10 in regulating IGF-I action is not known. The objective of this study was to determine the function of endogenous Grb10 in IGF signaling responses. Using small interfering RNA, we demonstrate that knockdown of Grb10 enhances IGF-I-mediated phosphorylation of insulin receptor substrate proteins, Akt/protein kinase B, and ERK1/2 and leads to a corresponding increase in DNA synthesis. Although IGF-I receptor autophosphorylation normally correlates with receptor signaling, we demonstrate a decrease in IGF-I-stimulated receptor phosphorylation in Grb10 knockdown cells. Pretreatment of cells with the protein-tyrosine phosphatase inhibitor pervanadate partially reverses this effect of Grb10 knockdown on receptor phosphorylation, indicating that endogenous Grb10 may block phosphatase access to the activated IGF-I receptor. Marked small interfering RNA knockdown of Grb10 does not result in increased or decreased expression of the related proteins Grb7 or Grb14. As further evidence for Grb10 functional specificity, the recently identified Grb10 interacting GYF proteins are shown to interact specifically with Grb10 and not with Grb7 or Grb14, using yeast two-hybrid assays. We conclude that Grb10 functions as a specific endogenous suppressor of IGF-I-stimulated cell signaling and DNA synthesis. Modulation of the Grb10-IGF-I receptor pathway may represent a mechanism that regulates IGF-I-responsive cell and tissue growth.
Dufresne
Aimee M
AM
Endocrinology Division and The Hallett Center for Diabetes and Endocrinology, Rhode Island Hospital, One Hoppin Street, Suite 200, Providence, Rhode Island 02903, USA.
Smith
Robert J
RJ
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004261:DNA Replication; D051383:GRB10 Adaptor Protein; D006706:Homeostasis; D007334:Insulin-Like Growth Factor I / Q000502:physiology*; D020935:MAP Kinase Signaling System / Q000502:physiology; D051379:Mice; D010766:Phosphorylation; D011506:Proteins / Q000502:physiology*; D034741:RNA, Small Interfering; D011972:Receptor, Insulin / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection
D017931:DNA Primers; C495959:Grb10 protein, mouse; D011506:Proteins; D034741:RNA, Small Interfering; D051383:GRB10 Adaptor Protein; D007334:Insulin-Like Growth Factor I; D011972:Receptor, Insulin