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PMID: 12748278 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

p27Kip1 inhibition of GRB2-SOS formation can regulate Ras activation.

Molecular and cellular biology ·Vol. 23 ·No. 11 ·2003-06-00 ·Pages 3735-52

Moeller SJ, Head ED, Sheaff RJ

Abstract

p27(Kip1) (p27) is often inappropriately downregulated in aggressive human cancers. Although p27 can inhibit cyclin-dependent kinases (CDKs), low p27 does not always correlate with increased CDK activity. Furthermore, cells derived from p27(-/-) mice respond to antimitogens, maintain restriction point control, and do not deregulate CDKs. Thus, disruption of a p27 function other than CDK inhibition may contribute to the disease state. A yeast two-hybrid screen identified growth factor receptor-bound protein 2 (GRB2) as a p27 binding partner. We now demonstrate that p27 can inhibit GRB2 function by blocking its association with the guanine nucleotide exchange factor SOS. Endogenous p27 is rapidly exported from the nucleus to the cytoplasm in response to mitogen stimulation, where it binds GRB2 concomitant with a decrease in GRB2-associated SOS. As predicted, mitogen-stimulated p27(-/-) cells maintained their GRB2-SOS complexes for significantly longer. The Ras/mitogen-activated protein kinase pathway does not appear to be deregulated in cells lacking p27 despite excess GRB2-SOS, suggesting that additional control mechanisms are present. A transient-transfection approach was employed to show that p27 can inhibit Ras activation by targeting GRB2 and further revealed that the CDK and GRB2 inhibitory functions of p27 are separable and distinct. Thus, p27 downregulation may compromise control of Ras, one of the most common oncogenic events in human cancer.

MeSH Terms
Adaptor Proteins, Signal Transducing Animals Cell Cycle Proteins/genetics,metabolism Cell Line Culture Media, Serum-Free Cyclin E/genetics,metabolism Cyclin-Dependent Kinase Inhibitor p27 Cyclin-Dependent Kinases/antagonists & inhibitors,metabolism Enzyme Activation Enzyme Inhibitors/metabolism Fatty Acids, Unsaturated/metabolism Fibroblasts/cytology,metabolism GRB2 Adaptor Protein Humans MAP Kinase Signaling System/physiology Macromolecular Substances Mice Mice, Knockout Mitogen-Activated Protein Kinase Kinases/genetics,metabolism Mitogens/metabolism Protein Binding Protein Transport/physiology Proteins/genetics,metabolism Recombinant Fusion Proteins/metabolism Son of Sevenless Proteins/metabolism Tumor Suppressor Proteins/genetics,metabolism Two-Hybrid System Techniques ras Proteins/metabolism
Chemicals
Adaptor Proteins, Signal Transducing Cdkn1b protein, mouse Cell Cycle Proteins Culture Media, Serum-Free Cyclin E Enzyme Inhibitors Fatty Acids, Unsaturated GRB2 Adaptor Protein GRB2 protein, human Grb2 protein, mouse Macromolecular Substances Mitogens Proteins Recombinant Fusion Proteins Son of Sevenless Proteins Tumor Suppressor Proteins Cyclin-Dependent Kinase Inhibitor p27 Cyclin-Dependent Kinases Mitogen-Activated Protein Kinase Kinases ras Proteins leptomycin B
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Moeller Stephanie J
University of Minnesota Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Head Elizabeth D
Sheaff Robert J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.23.11.3735-3752.2003
Published
2003-06-00
Pages
3735-52
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC155227
Subset
IM
Grants
NCI NIH HHS · CA-87267 · United States
External Links
PubMed source DOI: 10.1128/MCB.23.11.3735-3752.2003 PMC Full Text View journal details
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