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PMID: 7537849 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Phosphotyrosine-dependent interaction of SHC and insulin receptor substrate 1 with the NPEY motif of the insulin receptor via a novel non-SH2 domain.

Molecular and cellular biology ·Vol. 15 ·No. 5 ·1995-05-00 ·Pages 2500-8

Gustafson TA, He W, Craparo A, Schaub CD, O'Neill TJ

Abstract

The SHC proteins have been implicated in insulin receptor (IR) signaling. In this study, we used the sensitive two-hybrid assay of protein-protein interaction to demonstrate that SHC interacts directly with the IR. The interaction is mediated by SHC amino acids 1 to 238 and is therefore independent of the Src homology 2 domain. The interaction is dependent upon IR autophosphorylation, since the interaction is eliminated by mutation of the IR ATP-binding site. In addition, mutational analysis of the Asn-Pro-Glu-Tyr (NPEY) motif within the juxtamembrane domain of the IR showed the importance of the Asn, Pro, and Tyr residues to both SHC and IR substrate 1 (IRS-1) binding. We conclude that SHC interacts directly with the IR and that phosphorylation of Tyr-960 within the IR juxtamembrane domain is necessary for efficient interaction. This interaction is highly reminiscent of that of IRS-1 with the IR, and we show that the SHC IR-binding domain can substitute for that of IRS-1 in yeast and COS cells. We identify a homologous region within the IR-binding domains of SHC and IRS-1, which we term the SAIN (SHC and IRS-1 NPXY-binding) domain, which may explain the basis of these interactions. The SAIN domain appears to represent a novel motif which is able to interact with autophosphorylated receptors such as the IR.

MeSH Terms
Amino Acid Sequence Animals Binding Sites/genetics Cell Line Cloning, Molecular Humans Insulin Receptor Substrate Proteins Models, Biological Molecular Sequence Data Mutagenesis, Site-Directed Phosphoproteins/metabolism Phosphotyrosine Proteins/genetics,metabolism Receptor, Insulin/genetics,metabolism Saccharomyces cerevisiae/genetics Sequence Homology, Amino Acid Signal Transduction Tyrosine/analogs & derivatives,metabolism
Chemicals
IRS1 protein, human Insulin Receptor Substrate Proteins Phosphoproteins Proteins Phosphotyrosine Tyrosine Receptor, Insulin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gustafson T A
Department of Physiology, University of Maryland School of Medicine, Baltimore 21201, USA.
He W
Craparo A
Schaub C D
O'Neill T J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.5.2500
Published
1995-05-00
Pages
2500-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230480
Subset
IM
Grants
NIDDK NIH HHS · DK44093 · United States
NIGMS NIH HHS · GM08181 · United States
External Links
PubMed source DOI: 10.1128/MCB.15.5.2500 PMC Full Text View journal details
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