Reference Detail

Ref Type Journal Article
PMID (12370306)
Authors Delaney AM, Printen JA, Chen H, Fauman EB, Dudley DT
Title Identification of a novel mitogen-activated protein kinase kinase activation domain recognized by the inhibitor PD 184352.
Journal Molecular and cellular biology
Vol 22
Issue 21
Date 2002 Nov
URL
Abstract Text Utilizing a genetic screen in the yeast Saccharomyces cerevisiae, we identified a novel autoactivation region in mammalian MEK1 that is involved in binding the specific MEK inhibitor, PD 184352. The genetic screen is possible due to the homology between components of the yeast pheromone response pathway and the eukaryotic Raf-MEK-ERK signaling cascade. Using the FUS1::HIS3 reporter as a functional readout for activation of a reconstituted Raf-MEK-ERK signaling cascade, randomly mutagenized MEK variants that were insensitive to PD 184352 were obtained. Seven single-base-change mutations were identified, five of which mapped to kinase subdomains III and IV of MEK. Of the seven variants, only one, a leucine-to-proline substitution at amino acid 115 (Leu115Pro), was completely insensitive to PD 184352 in vitro (50% inhibitory concentration >10 micro M). However, all seven mutants displayed strikingly high basal activity compared to wild-type MEK. Overexpression of the MEK variants in HEK293T cells resulted in an increase in mitogen-activated protein (MAP) kinase phosphorylation, a finding consistent with the elevated basal activity of these constructs. Further, treatment with PD 184352 failed to inhibit Leu115Pro-stimulated MAP kinase activation in HEK293T cells, whereas all other variants had some reduction in phospho-MAP kinase levels. By using cyclic AMP-dependent protein kinase (1CDK) as a template, an MEK homology model was generated, with five of the seven identified residues clustered together, forming a potential hydrophobic binding pocket for PD 184352. Additionally, the model allowed identification of other potential residues that would interact with the inhibitor. Directed mutation of these residues supported this region's involvement with inhibitor binding.

Filtering

  • Case insensitive filtering will display rows where any text in any cell matches the filter term
  • Simple literal full or partial string matches
  • Separate multiple filter terms with a spaces, order doesn't matter (a b c and c b a are equivalent )
  • Filtering will only apply to rows that are already loaded on the page, filtering has no impact on query parameters
  • Use quotes to match a longer phrase which contains spaces "mtor c1483f"

Sorting

  • Generally, the default sort order for tables is set to be first column ascending, however, specific tables may set a different default sort order.
  • Click on any column header arrows to sort by that column
  • Hold down the Shift key and click multiple columns to sort by more than one column, be sure to set ascending or descending order for a given column, before moving on to the next column.

Molecular Profile Treatment Approach
Gene Name Source Synonyms Protein Domains Gene Description Gene Role
Therapy Name Therapy Description
Drug Name Trade Name Synonyms Drug Classes Drug Description
Variant Impact Protein Effect Variant Description Associated with drug Resistance
F53S missense unknown MAP2K1 F53S does not lie within any known functional domains of the Map2k1 protein (UniProt.org). The functional effect of F53S is conflicting, as F53S has been reported to confer a gain of function to the Map2k1 protein as indicated by Erk phosphorylation greater than that induced by wild-type, but not as active as other constitutively active Map2k1 mutants in culture (PMID: 16439621, PMID: 12370306) and in another study, F53S had autophosphorylation levels similar to wild-type Map2k1 (PMID: 29753091).
I103N missense gain of function - predicted MAP2K1 I103N lies within the protein kinase domain of the Map2k1 protein (UniProt.org). I103N is predicted to confer a gain of function on the Map2k1 protein as demonstrated by increased Map2k1 autophosphorylation in cell culture (PMID: 29753091) and is also associated with resistance to Mek inhibitors (PMID: 12370306, PMID: 19915144). Y
I111A missense gain of function - predicted MAP2K1 I111A lies within the protein kinase domain of the Map2k1 protein (UniProt.org). I111A is predicted to confer a gain of function to the Map2k1 protein, as demonstrated by increased Map2k1 kinase activity in an in-vitro assay and is also associated with resistance to inhibitors (PMID: 12370306). Y
I111N missense gain of function - predicted MAP2K1 I111N lies within the protein kinase domain of the Map2k1 protein (UniProt.org). I111N is predicted to confer a gain of function to the Map2k1 protein, as demonstrated by increased Map2k1 kinase activity in an in-vitro assay and is also associated with resistance to inhibitors (PMID: 12370306). Y
I111P missense gain of function - predicted MAP2K1 I111P lies within the protein kinase domain of the Map2k1 protein (UniProt.org). I111P is predicted to confer a gain of function to the Map2k1 protein, as demonstrated by increased Map2k1 kinase activity in an in-vitro assay and is also associated with resistance to inhibitors (PMID: 12370306). Y
I111R missense gain of function - predicted MAP2K1 I111R lies within the protein kinase domain of the Map2k1 protein (UniProt.org). I111R is predicted to confer a gain of function to the Map2k1 protein, as demonstrated by increased Map2k1 kinase activity in an in-vitro assay and is also associated with resistance to inhibitors (PMID: 12370306). Y
L115A missense gain of function - predicted MAP2K1 L115A lies within the protein kinase domain of the Map2k1 protein (UniProt.org). L115A is predicted to confer a gain of function to the Map2k1 protein, as demonstrated in an in-vitro kinase assay (PMID: 12370306) and is also associated with decreased binding and partial response to Mek inhibitors (PMID: 12370306).
L115P missense gain of function MAP2K1 L115P lies within the protein kinase domain of the Map2k1 protein (UniProt.org). L115P confers a gain of function on the kinase activity of the Map2k1 protein, as demonstrated by increased autophosphorylation (PMID: 29753091) and increased basal kinase activity (PMID: 12370306), and is also associated with decreased binding and resistance to Mek inhibitors (PMID: 12370306, PMID: 19915144, PMID: 26399658). Y
Molecular Profile Indication/Tumor Type Response Type Therapy Name Approval Status Evidence Type Efficacy Evidence References
MAP2K1 L115A Advanced Solid Tumor decreased response CI-1040 Preclinical - Cell culture Actionable In a preclinical study, CI-1040 (PD184352) partially inhibited kinase activity of Map2k1 L115A expressed in transformed human kidney cells (PMID: 12370306). 12370306
MAP2K1 F53S Advanced Solid Tumor sensitive CI-1040 Preclinical - Cell culture Actionable In a preclinical study, CI-1040 (PD184352) inhibited kinase activity of Map2k1 F53S expressed in transformed human kidney cells (PMID: 12370306). 12370306
MAP2K1 I103N Advanced Solid Tumor decreased response CI-1040 Preclinical - Cell culture Actionable In a preclinical study, CI-1040 (PD184352) partially inhibited kinase activity of Map2k1 I103N expressed in transformed human kidney cells (PMID: 12370306). 12370306
MAP2K1 L115P Advanced Solid Tumor resistant CI-1040 Preclinical - Cell culture Actionable In a preclinical study, CI-1040 (PD184352) did not inhibit kinase activity of Map2k1 L115P expressed in transformed human kidney cells (PMID: 12370306). 12370306