In planta changes in protein phosphorylation induced by the plant hormone abscisic acid.
Abscisic acid (ABA) is a hormone that controls seed dormancy and germination as well as the overall plant response to important environmental stresses such as drought. Recent studies have demonstrated that the ABA-bound receptor binds to and inhibits a class of protein phosphatases. To identify more broadly the phosphoproteins affected by this hormone in vivo, we used (14)N/(15)N metabolic labeling to perform a quantitative untargeted mass spectrometric analysis of the Arabidopsis thaliana phosphoproteome following ABA treatment. We found that 50 different phosphopeptides had their phosphorylation state significantly altered by ABA over a treatment period lasting 5-30 min. Among these changes were increases in phosphorylation of subfamily 2 SNF1-related kinases and ABA-responsive basic leucine zipper transcription factors implicated in ABA signaling by previous in vitro studies. Furthermore, four members of the aquaporin family showed decreased phosphorylation at a carboxy-terminal serine which is predicted to cause closure of the water-transporting aquaporin gate, consistent with ABA's role in ameliorating the effect of drought. Finally, more than 20 proteins not previously known to be involved with ABA were found to have significantly altered phosphorylation levels. Many of these changes are phosphorylation decreases, indicating that an expanded model of ABA signaling, beyond simple phosphatase inhibition, may be necessary. This quantitative proteomics dataset provides a more comprehensive, albeit incomplete, view both of the protein targets whose biochemical activities are likely to be controlled by ABA and of the nature of the emerging phosphorylation and dephosphorylation cascades triggered by this hormone.
Kline
Kelli G
KG
Biotechnology Center, Department of Biochemistry, University of Wisconsin, Madison, 53706, USA.
Barrett-Wilt
Gregory A
GA
Sussman
Michael R
MR
D000040:Abscisic Acid / Q000378:metabolism*; D000595:Amino Acid Sequence; D020346:Aquaporins / Q000737:chemistry / Q000378:metabolism; D017360:Arabidopsis / Q000378:metabolism; D017124:Conserved Sequence; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010937:Plant Growth Regulators / Q000378:metabolism*; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*
D020346:Aquaporins; D010937:Plant Growth Regulators; D010940:Plant Proteins; D000040:Abscisic Acid