Regulation of wound-responsive calcium-dependent protein kinase from maize (ZmCPK11) by phosphatidic acid

• Published in: Acta biochimica Polonica Vol. 58; no. 4; p. 589
• Main Authors: Klimecka, Maria, Szczegielniak, Jadwiga, Godecka, Luiza, Lewandowska-Gnatowska, Elżbieta, Dobrowolska, Grażyna, Muszyńska, Grażyna
• Format: Journal Article
• Language: English
• Published: Poland 01-01-2011
• Subjects: 1-Butanol - pharmacology; Enzyme Activation; Enzyme Assays; Gene Expression Regulation, Plant; Genes, Plant; Mutagenesis, Site-Directed; Phosphatidic Acids - metabolism; Phosphatidic Acids - pharmacology; Phospholipase D - genetics; Phospholipase D - metabolism; Phosphorylation; Plant Leaves - drug effects; Plant Leaves - genetics; Plant Leaves - metabolism; Plant Proteins - genetics; Plant Proteins - metabolism; Protein Binding; Protein Kinases - genetics; Protein Kinases - metabolism; Protoplasts - cytology; Protoplasts - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; RNA, Messenger - genetics; RNA, Messenger - metabolism; Signal Transduction; Stress, Physiological; Time Factors; Transfection; Zea mays - drug effects; Zea mays - enzymology; Zea mays - genetics
• ISSN: 0001-527X; 1734-154X
• DOI: 10.18388/abp.2011_2229

In plant cells, phospholipids are not only membrane components but also act as second messengers interacting with various proteins and regulating diverse cellular processes, including stress signal transduction. Here, we report studies on the effects of various phospholipids on the activity and expression of maize wound-responsive calcium-dependent protein kinase (ZmCPK11). Our results revealed that in leaves treated with n-butanol, a potent inhibitor of phosphatidic acid (PA) synthesis catalyzed by phospholipase D, a significant decrease of ZmCPK11 activity was observed, indicating contribution of PA in the kinase activation. Using lipid binding assays, we demonstrate that among various phospholipids only saturated acyl species (16:0 and 18:0) of phosphatidic acid are able to bind to ZmCPK11. Saturated acyl species of PA are also able to stimulate phosphorylation of exogenous substrates by ZmCPK11 and autophosphorylation of the kinase. The level of ZmCPK11 autophosphorylation is correlated with its enzymatic activity. RT-PCR analysis showed that transcript level of ZmCPK11 in maize leaves increased in response to PA treatment. The influence of PA on the activity and transcript level of ZmCPK11 suggests an involvement of this kinase in a PA-mediated wound signal transduction pathway.