Membrane localization of a rice calcium-dependent protein kinase (CDPK) is mediated by myristoylation and palmitoylation.
Calcium-dependent protein kinases (CDPKs), the most abundant serine/threonine kinases in plants, are found in various subcellular localizations, which suggests that this family of kinases may be involved in multiple signal transduction pathways. A complete analysis to try to understand the molecular basis of the presence of CDPKs in various localizations in the cell has not been accomplished yet. It has been suggested that myristoylation may be responsible for membrane association of CDPKs. In this study, we used a rice CDPK, OSCPK2, which has a consensus sequence for myristoylation at the N-terminus, to address this question. We expressed wild-type OSCPK2 and various mutants in different heterologous systems to investigate the factors that affect its membrane association. The results show that OSCPK2 is myristoylated and palmitoylated and targeted to the membrane fraction. Both modifications are required, myristoylation being essential for membrane localization and palmitoylation for its full association. The fact that palmitoylation is a reversible modification may provide a mechanism for regulation of the subcellular localization. OSCPK2 is the first CDPK shown to be targeted to membranes by an src homology domain 4 (SH4) located at the N-terminus of the molecule.
Martín
M L
ML
Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Centro de Investigaciones Biológicas, Fundación para Investigaciones Biológicas Aplicadas, Mar del Plata, Argentina.
Busconi
L
L
D001692:Biological Transport; D002118:Calcium / Q000494:pharmacology; D004274:DNA, Recombinant / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008566:Membranes / Q000201:enzymology*; D009154:Mutation; D019814:Myristic Acid / Q000378:metabolism*; D012275:Oryza / Q000201:enzymology*; D010169:Palmitic Acids / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011523:Protoplasts / Q000378:metabolism; D003313:Zea mays / Q000235:genetics
D004274:DNA, Recombinant; D010169:Palmitic Acids; D019814:Myristic Acid; D017346:Protein Serine-Threonine Kinases; D002118:Calcium