Negative feedback regulation of microbe-associated molecular pattern-induced cytosolic Ca2+ transients by protein phosphorylation
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Microbe/pathogen-associated molecular patterns (MAMPs/PAMPs) often induce rises in cytosolic free Ca2+ concentration ([Ca2+]cyt) and protein phosphorylation. Though they are postulated to play pivotal roles in plant innate immunity, their molecular links and the regulatory mechanisms remain largely unknown. To investigate the regulatory mechanisms for MAMP-induced Ca2+ mobilization, we have established a transgenic rice (Oryza sativa) cell line stably expressing apoaequorin, and characterized the interrelationship among MAMP-induced changes in [Ca2+]cyt, production of reactive oxygen species (ROS) and protein phosphorylation. Oligosaccharide and sphingolipid MAMPs induced Ca2+ transients mainly due to plasma membrane Ca2+ influx, which were dramatically suppressed by a protein phosphatase inhibitor, calyculin A (CA). Hydrogen peroxide and hypo-osmotic shock triggered similar [Ca2+]cyt elevations, which were not affected by CA. MAMP-induced protein phosphorylation, which is promoted by CA, has been shown to be required for ROS production and MAPK activation, while it negatively regulates MAMPs-induced Ca2+ mobilization and may play a crucial role in temporal regulation of [Ca2+]cyt signature.
KeywordsCalcium ion Calyculin A Microbe/pathogen-associated molecular patterns (MAMPs/PAMPs) Protein phosphorylation Reactive oxygen species
We would like to thank Prof. Dr. Naoto Shibuya of Meiji University for gift of N-cetylchitooligosaccharides and suggestions, Prof. Dr. Kenzo Nakamura for the pIG121-Hm plasmid, Prof. Dr. Ko Shimamoto for suggestions, and Mr. Jumpei Hamada, Mr. Yohei Iwasaki, and Mr. Hiroshi Nokajima for technical assistance. This work was supported in part by Grant-in-Aid for Scientific Research on Innovative Areas (21200067 to T.K.), for Exploratory Research (21658118 to K.K.) and for Young Scientists (B) (21780041 to T.K.), and by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Integrated research project for plants, insects, and animals using genome technology MP-2134 to K.K.).
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