Linked Life Data

9553107

Source:http://linkedlifedata.com/resource/pubmed/id/9553107

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
17
pubmed:dateCreated
1998-5-22
pubmed:databankReference
pubmed:abstractText
Grb10 and its close homologues Grb7 and Grb14, belong to a family of adapter proteins characterized by a proline-rich region, a central PH domain, and a carboxyl-terminal Src homology 2 (SH2) domain. Their interaction with a variety of activated tyrosine kinase receptors is well documented, but their actual function remains a mystery. The Grb10 SH2 domain was isolated from a two-hybrid screen using the MEK1 kinase as a bait. We show that this unusual SH2 domain interacts, in a phosphotyrosine-independent manner, with both the Raf1 and MEK1 kinases. Mutation of the MEK1 Thr-386 residue, which is phosphorylated by mitogen-activated protein kinase in vitro, reduces binding to Grb10 in a two-hybrid assay. Interaction of Grb10 with Raf1 is constitutive, while interaction between Grb10 and MEK1 needs insulin treatment of the cells and follows mitogen-activated protein kinase activation. Random mutagenesis of the SH2 domain demonstrated that the Arg-betaB5 and Asp-EF2 residues are necessary for binding to the epidermal growth factor and insulin receptors as well as to the two kinases. In addition, we show that a mutation in Ser-betaB7 affects binding only to the receptors, while a mutation in Thr-betaC5 abrogates binding only to MEK1. Finally, transfection of Grb10 genes with specific mutations in their SH2 domains induces apoptosis in HTC-IR and COS-7 cells. These effects can be competed by co-expression of the wild type protein, suggesting that these mutants act by sequestering necessary signaling components.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
24
pubmed:volume
273
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
10475-84
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:9553107-Amino Acid Sequence, pubmed-meshheading:9553107-Animals, pubmed-meshheading:9553107-Apoptosis, pubmed-meshheading:9553107-Base Sequence, pubmed-meshheading:9553107-Binding Sites, pubmed-meshheading:9553107-Cell Line, pubmed-meshheading:9553107-DNA, pubmed-meshheading:9553107-GRB10 Adaptor Protein, pubmed-meshheading:9553107-Humans, pubmed-meshheading:9553107-MAP Kinase Kinase 1, pubmed-meshheading:9553107-Mitogen-Activated Protein Kinase Kinases, pubmed-meshheading:9553107-Molecular Sequence Data, pubmed-meshheading:9553107-Phosphorylation, pubmed-meshheading:9553107-Point Mutation, pubmed-meshheading:9553107-Protein Binding, pubmed-meshheading:9553107-Protein-Serine-Threonine Kinases, pubmed-meshheading:9553107-Protein-Tyrosine Kinases, pubmed-meshheading:9553107-Proteins, pubmed-meshheading:9553107-Proto-Oncogene Proteins c-raf, pubmed-meshheading:9553107-Sequence Homology, Amino Acid, pubmed-meshheading:9553107-Sequence Homology, Nucleic Acid
pubmed:year
1998
pubmed:articleTitle
Interaction of the Grb10 adapter protein with the Raf1 and MEK1 kinases.
pubmed:affiliation
Eukaryotic Genetics Group, Biotechnology Research Institute, National Research Council, 6100 Royalmount, Montreal, Quebec H4P 2R2, Canada. andre.nantel@bri.nrc.ca
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't