Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Analysis of Dual Phosphorylation of Hog1 MAP Kinase in Saccharomyces cerevisiae Using Quantitative Mass Spectrometry

  • Choi, Min-Yeon (Department of Biological Sciences, Seoul National University) ;
  • Kang, Gum-Yong (Department of Molecular Biotechnology, Konkuk University) ;
  • Hur, Jae-Young (Department of Biological Sciences, Seoul National University) ;
  • Jung, Jin Woo (Department of Molecular Biotechnology, Konkuk University) ;
  • Kim, Kwang Pyo (Department of Molecular Biotechnology, Konkuk University) ;
  • Park, Sang-Hyun (Department of Biological Sciences, Seoul National University)
  • Received : 2008.04.02
  • Accepted : 2008.04.14
  • Published : 2008.08.31
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Abstract

The mitogen-activated protein kinase (MAPK) signaling pathway is activated in response to extracellular stimuli and regulates various activities in eukaryotic cells. Following exposure to stimuli, MAPK is known to be activated via dual phosphorylation at a conserved TxY motif in the activation loop; both threonine and tyrosine residues are phosphorylated by an upstream kinase. However, the mechanism underlying dual phosphorylation is not clearly understood. In the budding yeast Saccharomyces cerevisiae, the Hog1 MAPK mediates the high-osmolarity glycerol (HOG) signaling pathway. Tandem mass spectrometry and phosphospecific immunoblotting were performed to quantitatively monitor the dynamic changes occurring in the phosphorylation status of the TxY motif of Hog1 on exposure to osmotic stress. The results of our study suggest that the tyrosine residue is preferentially and dynamically phosphorylated following stimulation, and this in turn leads to the dual phosphorylation. The tyrosine residue was hyperphosphorylated in the absence of a threonine residue; this result suggests that the threonine residue is critical for the control of signaling noise and adaptation to osmotic stress.

Keywords

Acknowledgement

Supported by : Korea Research Foundation

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