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PMID: 16043515 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The p85 regulatory subunit of phosphoinositide 3-kinase down-regulates IRS-1 signaling via the formation of a sequestration complex.

The Journal of cell biology ·Vol. 170 ·No. 3 ·2005-08-01 ·Pages 455-64

Luo J, Field SJ, Lee JY, Engelman JA, Cantley LC

Abstract

Phosphoinositide (PI) 3-kinase is required for most insulin and insulin-like growth factor (IGF) 1-dependent cellular responses. The p85 regulatory subunit of PI 3-kinase is required to mediate the insulin-dependent recruitment of PI 3-kinase to the plasma membrane, yet mice with reduced p85 expression have increased insulin sensitivity. To further understand the role of p85, we examined IGF-1-dependent translocation of p85alpha by using a green fluorescence protein (GFP)-tagged p85alpha (EGFP-p85alpha). In response to IGF-1, but not to PDGF signaling, EGFP-p85alpha translocates to discrete foci in the cell. These foci contain the insulin receptor substrate (IRS) 1 adaptor molecule, and their formation requires the binding of p85 to IRS-1. Surprisingly, monomeric p85 is preferentially localized to these foci compared with the p85-p110 dimer, and these foci are not sites of phosphatidylinositol-3,4,5-trisphosphate production. Ultrastructural analysis reveals that p85-IRS-1 foci are cytosolic protein complexes devoid of membrane. These results suggest a mechanism of signal down-regulation of IRS-1 that is mediated by monomeric p85 through the formation of a sequestration complex between p85 and IRS-1.

MeSH Terms
Animals CHO Cells Cricetinae Cricetulus Cytosol/metabolism Dimerization Down-Regulation Green Fluorescent Proteins/genetics Humans Insulin Receptor Substrate Proteins Mice Phosphatidylinositol 3-Kinases/genetics,metabolism,physiology Phosphoproteins/metabolism,physiology Phosphorylation Protein Binding Protein Subunits/metabolism Protein Transport Receptor, IGF Type 1/agonists Recombinant Fusion Proteins/metabolism Signal Transduction Tyrosine/metabolism
Chemicals
IRS1 protein, human Insulin Receptor Substrate Proteins Irs1 protein, mouse Phosphoproteins Protein Subunits Recombinant Fusion Proteins enhanced green fluorescent protein Green Fluorescent Proteins Tyrosine Phosphatidylinositol 3-Kinases Receptor, IGF Type 1
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Luo Ji
Department of Systems Biology, Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA 02115, USA.
Field Seth J
Lee Jennifer Y
Engelman Jeffrey A
Cantley Lewis C
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200503088
Published
2005-08-01
Epub
2005-00-25
Pages
455-64
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2171479
Subset
IM
Grants
NIDDK NIH HHS · K08 DK065108-01 · United States
NIGMS NIH HHS · R37 GM041890 · United States
NCI NIH HHS · CA089021 · United States
NIGMS NIH HHS · R01 GM041890 · United States
NCI NIH HHS · P01 CA089021 · United States
NIGMS NIH HHS · GM41890 · United States
NIDDK NIH HHS · K08 DK065108 · United States
External Links
PubMed source DOI: 10.1083/jcb.200503088 PMC Full Text View journal details
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