53BP2S, interacting with insulin receptor substrates, modulates insulin signaling

Fumihiko Hakuno, Shigekazu Kurihara, Robert T. Watson, Jeffrey E. Pessin, Shin Ichiro Takahashi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

It is well known that insulin receptor substrates (IRS) act as a mediator for signal transduction of insulin, insulin-like growth factors, and several cytokines. To identify proteins that interact with IRS and modulate IRS-mediated signals, we performed yeast two-hybrid screening with IRS-1 as bait. Out of 109 cDNA-positive clones identified from a human placental cDNA library, two clones encoded 53BP2, p53-binding protein 2 (53BP2S), a short form splicing variant of the apoptosis-stimulating protein of p53 that possesses Src homology region 3 domain, and ankyrin repeats domain, and had been reported to interact with p53, Bcl-2, and NF-κB. Interaction of 53BP2S with IRS-1 was confirmed by glutathione S-transferase pull-down and co-immunoprecipitation assays in COS-7 cells and 3T3-L1 adipocytes. The Src homology region 3 domain and ankyrin repeats domain of 53BP2S were responsible for its interaction with IRS-1, whereas the phosphotyrosine binding domain and a central domain (amino acid residues 750-861) of IRS-1 were required for its interaction with 53BP2S. In CHO-C400 cells, expression of 53BP2S reduced insulin-stimulated IRS-1 tyrosine phosphorylation with a concomitant enhancement of IRS-2 tyrosine phosphorylation. In addition, the amount of the phosphatidylinositol 3-kinase regulatory p85 subunit associated with tyrosine-phosphorylated proteins, and activation of Akt was enhanced by 53BP2S expression. Although 53BP2S also enhanced Akt activation in 3T3-L1 adipocytes, insulin-induced glucose transporter 4 translocation was markedly inhibited in accordance with reduction of insulin-induced AS160 phosphorylation. Together these data demonstrate that 53BP2S interacts and modulates the insulin signals mediated by IRSs.

Original languageEnglish (US)
Pages (from-to)37747-37758
Number of pages12
JournalJournal of Biological Chemistry
Volume282
Issue number52
DOIs
StatePublished - Dec 28 2007
Externally publishedYes

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Insulin Receptor Substrate Proteins
Insulin Receptor
Insulin
Phosphorylation
Substrates
Ankyrin Repeat
Adipocytes
Tyrosine
Clone Cells
Chemical activation
Phosphatidylinositol 3-Kinase
Signal transduction
Phosphotyrosine
Proteins
Facilitative Glucose Transport Proteins
CHO Cells
COS Cells
Somatomedins
Glutathione Transferase
Gene Library

ASJC Scopus subject areas

  • Biochemistry

Cite this

53BP2S, interacting with insulin receptor substrates, modulates insulin signaling. / Hakuno, Fumihiko; Kurihara, Shigekazu; Watson, Robert T.; Pessin, Jeffrey E.; Takahashi, Shin Ichiro.

In: Journal of Biological Chemistry, Vol. 282, No. 52, 28.12.2007, p. 37747-37758.

Research output: Contribution to journalArticle

Hakuno, Fumihiko ; Kurihara, Shigekazu ; Watson, Robert T. ; Pessin, Jeffrey E. ; Takahashi, Shin Ichiro. / 53BP2S, interacting with insulin receptor substrates, modulates insulin signaling. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 52. pp. 37747-37758.
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