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Volume 272, Number 18, Issue of May 2, 1997 pp. 11968-11974
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

CSF-1 Receptor/Insulin Receptor Chimera Permits CSF-1-dependent Differentiation of 3T3-L1 Preadipocytes

(Received for publication, January 17, 1997, and in revised form, February 21, 1997)

Oleg V. Chaika Dagger , Nina Chaika Dagger , Deanna J. Volle Dagger , Peter A. Wilden § , Samuel J. Pirrucello and Robert E. Lewis Dagger par

From the Dagger  Eppley Institute for Research in Cancer and Allied Diseases, Departments of par  Biochemistry and Molecular Biology and  Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska 68198-6805 and § Department of Pharmacology, University of Missouri School of Medicine, Columbia, Missouri 65212

A chimeric growth factor receptor (CSF1R/IR) was constructed by splicing cDNA sequences encoding the extracellular ligand binding domain of the human colony stimulating factor-1 (CSF-1) receptor to sequences encoding the transmembrane and cytoplasmic domains of the human insulin receptor. The addition of CSF-1 to cells transfected with the CSF1R/IR chimera cDNA stimulated the tyrosine phosphorylation of a protein that was immunoprecipitated by an antibody directed against the carboxyl terminus of the insulin receptor. Phosphopeptide maps of the 32P-labeled CSF1R/IR protein revealed the same pattern of phosphorylation observed in 32P-labeled insulin receptor beta  subunits. CSF-1 stimulated the tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and Shc in cells expressing the CSF1R/IR chimera. Lipid accumulation and the expression of a differentiation-specific marker demonstrated that 3T3-L1 preadipocytes undergo CSF-1-dependent differentiation when transfected with the CSF1R/IR chimera cDNA but not when transfected with the expression vector alone. A 12-amino acid deletion within the juxtamembrane region of the CSF1R/IR (CSF1R/IRDelta 960) blocked CSF-1-stimulated phosphorylation of IRS-1 and Shc but did not inhibit CSF-1-mediated differentiation of 3T3-L1 preadipocytes. These observations indicate that adipocyte differentiation can be initiated by intracellular pathways that do not require tyrosine phosphorylation of IRS-1 or Shc.


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Copyright © 1997 by the American Society for Biochemistry and Molecular Biology.