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Quantitative and functional post-translational modification (PTM) proteomics and interactomics have emerged as powerful
omics approaches in studying cellular events in various model organisms. In this seminar, I intend to show several examples on
how to apply in planta metabolic labeling and in vitro chemical labeling-based (4C) quantitative PTM proteomic and interactomic
workflow (SILIA, SQUA-D and AQUIP) in investigation of cell signaling in the model plant Arabidopsis and its potential impact in
the plant cell biology research in general. To elucidate the molecular mechanism underlying plant hormone ethylene signaling in
Arabidopsis on a number of plant responses, several well-known Arabidopsis ethylene response loss-of-function mutants (ctr1-1,
rcn1-1, ein2-5 and eil3eil1) were selected as target plant materials for both stable isotope metabolic labeling (SILIA) and in vitro
dimethyl labeling (SQUA-D) for the PTM quantitation. The 4C quantitative proteomics and interactomics results clearly revealed that
there exist multiple PTM-mediated signaling pathways in Arabidopsis. This quantitative PTM proteomics was able to identify rapidly
phosphorylated proteins, such as TREPH1, MAP Kinase Kinases, CPKs, in response to 40 second of touch or 150 seconds of gravity
stimulation in Arabidopsis. The following reverse genetic and transgenic plant approaches in combination with cell biology studies
validated the biological functions of these key candidate phosphoproteins in these internal and external signals-mediated cellular
events and dramatic plant responses. These successful research results suggest that our PTM proteomic approach can be quantitative,
repeatable, accurate and versatile in addressing many important biological questions in life sciences.