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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
23
|
| pubmed:dateCreated |
1993-9-15
|
| pubmed:abstractText |
Treatment of cells with various growth factors and mitogens results in the rapid hyperphosphorylation and activation of the Raf-1 kinase. To determine if phosphorylation events affect Raf-1 activity, we have initiated experiments to identify the phosphorylation sites of Raf-1. In this report, we find that Ser43, Ser259, and Ser621 are the major sites of Raf-1 which are phosphorylated in mammalian cells and in Sf9 insect cells infected with a recombinant baculovirus encoding human Raf-1. Mutant Raf-1 proteins lacking kinase activity are also phosphorylated on these sites in vivo, indicating that these phosphorylation events are not a consequence of autophosphorylation. Furthermore, we find that Thr268 is the predominant Raf-1 residue phosphorylated in in vitro autokinase assays. In addition, we have examined the biochemical activity of baculovirus-expressed Raf-1 proteins containing mutations at these phosphorylation sites. In in vitro protein kinase assays Ser259 mutant proteins were 2-fold more active than wild-type Raf-1 and Ser621 mutant proteins were inactive as kinases. Analysis of the residues surrounding Ser259 and Ser621 indicates that RSXSXP may be a consensus sequence for the kinase responsible for phosphorylation of Raf-1 at these sites. Interestingly, these RSXSXP sequences are completely conserved throughout evolution in all Raf family members.
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| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Protein-Serine-Threonine Kinases,
http://linkedlifedata.com/resource/pubmed/chemical/Proto-Oncogene Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Proto-Oncogene Proteins c-raf,
http://linkedlifedata.com/resource/pubmed/chemical/Serine
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Aug
|
| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
15
|
| pubmed:volume |
268
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
17309-16
|
| pubmed:dateRevised |
2009-11-19
|
| pubmed:meshHeading |
pubmed-meshheading:8349614-3T3 Cells,
pubmed-meshheading:8349614-Amino Acid Sequence,
pubmed-meshheading:8349614-Animals,
pubmed-meshheading:8349614-Baculoviridae,
pubmed-meshheading:8349614-Cell Line,
pubmed-meshheading:8349614-Chromatography, High Pressure Liquid,
pubmed-meshheading:8349614-Cloning, Molecular,
pubmed-meshheading:8349614-Electrophoresis, Gel, Two-Dimensional,
pubmed-meshheading:8349614-Humans,
pubmed-meshheading:8349614-Mice,
pubmed-meshheading:8349614-Molecular Sequence Data,
pubmed-meshheading:8349614-Moths,
pubmed-meshheading:8349614-Mutagenesis, Site-Directed,
pubmed-meshheading:8349614-Phosphorylation,
pubmed-meshheading:8349614-Protein-Serine-Threonine Kinases,
pubmed-meshheading:8349614-Proto-Oncogene Proteins,
pubmed-meshheading:8349614-Proto-Oncogene Proteins c-raf,
pubmed-meshheading:8349614-Rats,
pubmed-meshheading:8349614-Serine
|
| pubmed:year |
1993
|
| pubmed:articleTitle |
Identification of the major phosphorylation sites of the Raf-1 kinase.
|
| pubmed:affiliation |
ABL-Basic Research Program, National Cancer Institute-Frederick Cancer Research and Development Center, Maryland 21702.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|