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Gene/Protein
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Target Concepts:
Gene/Protein
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Query: EC:3.4.11.18 (
MAP
)
7,412
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
The Arabidopsis MAP kinase 4 (MPK4) substrate
MKS1
was expressed in Escherichia coli and purified, full-length, 6x histidine (His)-tagged
MKS1
was phosphorylated in vitro by hemagglutinin (HA)-tagged MPK4 immuno-precipitated from plants.
MKS1
phosphorylation was initially verified by electrophoresis and gel-staining with ProQ Diamond and the protein was digested by either trypsin or chymotrypsin for maximum sequence coverage to facilitate identification of phosphorylated positions. Prior to analysis by mass spectrometry, samples were either desalted, passed over TiO(2) or both for improved phosphopeptide detection. As
MAP
kinases generally phosphorylate serine or threonine followed by proline (Ser/Thr-Pro), theoretical masses of potentially phosphorylated peptides were calculated and mass spectrometric peaks matching these masses were fragmented and searched for a neutral-loss signal at approximately 98 Da indicative of phosphorylation. Additionally, mass spectrometric peaks present in the MPK4-treated
MKS1
, but not in the control peptide map of untreated
MKS1
, were fragmented. Fragmentation spectra were subjected to a MASCOT database search which identified three of the twelve Ser-Pro serine residues (Ser72, Ser108, Ser120) in the phosphorylated form.
...
PMID:Phosphorylation sites of Arabidopsis MAP kinase substrate 1 (MKS1). 1770 78
Plant and animal perception of microbes through pathogen surveillance proteins leads to MAP kinase signalling and the expression of defence genes. However, little is known about how plant
MAP
kinases regulate specific gene expression. We report that, in the absence of pathogens, Arabidopsis MAP kinase 4 (MPK4) exists in nuclear complexes with the WRKY33 transcription factor. This complex depends on the MPK4 substrate
MKS1
. Challenge with Pseudomonas syringae or flagellin leads to the activation of MPK4 and phosphorylation of
MKS1
. Subsequently, complexes with
MKS1
and WRKY33 are released from MPK4, and WRKY33 targets the promoter of PHYTOALEXIN DEFICIENT3 (PAD3) encoding an enzyme required for the synthesis of antimicrobial camalexin. Hence, wrky33 mutants are impaired in the accumulation of PAD3 mRNA and camalexin production upon infection. That WRKY33 is an effector of MPK4 is further supported by the suppression of PAD3 expression in mpk4-wrky33 double mutant backgrounds. Our data establish direct links between MPK4 and innate immunity and provide an example of how a plant MAP kinase can regulate gene expression by releasing transcription factors in the nucleus upon activation.
...
PMID:Arabidopsis MAP kinase 4 regulates gene expression through transcription factor release in the nucleus. 1865 Sep 34