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Cloning of a cDNA encoding a 190-kDa insulin receptor substrate-1-like protein of simian COS cells

L Wang1, H Hayashi, Y Mitani

  • 1Department of Enzyme Genetics, University of Tokushima, Japan.

Biochemical and Biophysical Research Communications
|November 2, 1995
PubMed

Insights

Researchers identified a novel insulin receptor substrate-like protein (pp190) in simian cells, closely resembling human insulin receptor substrate-1 (IRS-1). This finding suggests a conserved role for IRS-1 in insulin signaling across species.

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Biochemistry

Background:

  • Insulin receptor substrate-1 (IRS-1) is crucial for mediating major insulin signals, including glucose uptake and DNA synthesis.
  • IRS-1 phosphorylation is a key step in insulin signal transduction pathways.

Purpose of the Study:

  • To clone and characterize a novel IRS-1-like protein from simian COS cells.
  • To investigate the relationship between this novel protein and known IRS proteins (IRS-1 and IRS-2).

Main Methods:

  • Cloning of cDNA encoding a 190-kDa IRS-1-like protein (pp190) from simian COS cells.
  • Deduced amino acid sequence analysis and comparison with human, mouse, and rat IRS-1.
  • Binding assays to assess interactions with SH2 domain-containing proteins (p85, Grb2/Ash, SH-PTP2).
  • Southern blotting to investigate gene presence in Chinese hamster ovary (CHO) cells.

Main Results:

  • A 190-kDa IRS-1-like protein (pp190) was cloned from COS cells, showing high amino acid identity (96.4%) to human IRS-1.
  • COS pp190 demonstrated binding to SH2 domains of p85, Grb2/Ash, and SH-PTP2, similar to IRS-1.
  • Southern blotting indicated that CHO cells possess both the IRS-1 gene and a gene related to COS pp190.

Conclusions:

  • The cloned COS pp190 is likely a simian homologue of IRS-1, not IRS-2, suggesting evolutionary conservation of IRS-1 function.
  • The presence of related genes in CHO cells hints at a broader gene family involved in insulin signaling.

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