Reexamining interaction of the SH2 domains of SYP and GAP with insulin and IGF-1 receptors in the two-hybrid system.
Direct interaction of effector proteins such as the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI 3-kinase), SYP (SH2-domain-containing tyrosine phosphatase) and GAP (Ras-GTPase activating protein) with the insulin receptor (IR) and insulin-like growth factor-1 (IGF-1) type 1 receptor (IGF-1R) has been reported in some studies. Interaction of SYP and GAP with IR and IGF-1R was re-investigated here in the two-hybrid system by assessing his3/lacZ activation in S. cerevisiae. The experiments were performed with the cytoplasmic beta domain of IR and IGF-1R and various SH2-subdomains of SYP and GAP. None of the subdomains of SYP and GAP tested were able to activate his3/lacZ, whereas these reporter genes were strongly activated when p85 was used as we have recently shown. Thus, interaction of SYP and GAP with IR and IGF-1R, if any, would be weak and/or transient as compared to that of p85.
Lamothe
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B
Institut Cochin de Génétique Moléculaire, INSERM U257, Paris, France.
Bucchini
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Jami
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Joshi
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D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002455:Cell Division / Q000235:genetics; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D020690:GTPase-Activating Proteins; D017930:Genes, Reporter / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D017526:Receptor, IGF Type 1 / Q000737:chemistry / Q000378:metabolism*; D011972:Receptor, Insulin / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D015533:Transcriptional Activation / Q000235:genetics; D020703:ras GTPase-Activating Proteins; D018909:src Homology Domains / Q000235:genetics*; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D020690:GTPase-Activating Proteins; D047908:Intracellular Signaling Peptides and Proteins; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases; D011506:Proteins; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin; D011993:Recombinant Fusion Proteins; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases; D014157:Transcription Factors; D020703:ras GTPase-Activating Proteins; D000099160:LexA Repressor Protein; C052130:GAL4 protein, S cerevisiae