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1.
Mol Cells ; 42(9): 646-660, 2019 Sep 30.
Article in English | MEDLINE | ID: mdl-31480825

ABSTRACT

Abscisic acid (ABA) is a phytohormone essential for seed development and seedling growth under unfavorable environmental conditions. The signaling pathway leading to ABA response has been established, but relatively little is known about the functional regulation of the constituent signaling components. Here, we present several lines of evidence that Arabidopsis Raf-like kinase Raf10 modulates the core ABA signaling downstream of signal perception step. In particular, Raf10 phosphorylates subclass III SnRK2s (SnRK2.2, SnRK2.3, and SnRK2.6), which are key positive regulators, and our study focused on SnRK2.3 indicates that Raf10 enhances its kinase activity and may facilitate its release from negative regulators. Raf10 also phosphorylates transcription factors (ABI5, ABF2, and ABI3) critical for ABAregulted gene expression. Furthermore, Raf10 was found to be essential for the in vivo functions of SnRK2s and ABI5. Collectively, our data demonstrate that Raf10 is a novel regulatory component of core ABA signaling.


Subject(s)
Abscisic Acid/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , MAP Kinase Kinase Kinases/metabolism , Signal Transduction , Amino Acid Sequence , Arabidopsis Proteins/chemistry , MAP Kinase Kinase Kinases/chemistry , Phenotype , Phosphorylation , Phosphoserine/metabolism , Phosphothreonine/metabolism , Protein Multimerization
2.
Proteomics Clin Appl ; 13(5): e1900028, 2019 09.
Article in English | MEDLINE | ID: mdl-31119868

ABSTRACT

PURPOSE: Lung cancer is among the most common cancers. Bronchoalveolar lavage fluid (BALF) can be easily obtained from patients with lung cancers. The aim is to develop a novel proteomic method of the molecule-based sensitive detection of biomarkers from BALF. EXPERIMENTAL DESIGN: BALF samples are collected from segmental bronchus of 14 patients with lung cancers from Kyung Hee University Hospital. First, BALF proteome is depleted using a depletion column, and then peptides are prepared from the enriched low abundant proteins and fractionated by high pH reverse phase liquid chromatography prior to LC-MS/MS. Data are available via ProteomeXchange with identifier PXD012645. RESULTS: A novel method is developed for in-depth proteomic analysis of BALF by combining antibody-based depletion of high abundant proteins from BALF with high pH peptide fractionation. Peptides are analyzed on a high resolution Orbitrap Fusion mass spectrometer. MaxQuant search result in the identification of 4615 protein groups mapped to 4534 genes. CONCLUSIONS AND CLINICAL RELEVANCE: It is found that the method outperformed conventional BALF proteomic methods and it is believed that this method will facilitate the biomarker research for lung cancer. In addition, it is shown that BALF will be a great source of biomarkers of lung diseases.


Subject(s)
Biomarkers, Tumor/metabolism , Bronchoalveolar Lavage Fluid , Lung Neoplasms/metabolism , Proteomics , Humans
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