title: Sucrose induces rapid activation of CfSAPK, a mitogen-activated protein kinase, in Cephalostachyum fuchsianum Gamble cells.
abstract: Sucrose was recently demonstrated to function as a molecular signal. However, sucrose-specific sensing and signalling pathways remain largely undefined. Here, we show that Cephalostachyum fuchsianum sucrose-activated protein kinase (CfSAPK) is transiently and specifically activated by sucrose in C. fuchsianum Gamble suspension cells. The result suggested that CfSAPK participates in a sucrose-signalling pathway. CfSAPK was partially purified from sucrose-treated cells and further analysed. Kinase activity assays revealed that CfSAPK preferentially used myelin basic protein (MBP) as substrate in vitro and strongly phosphorylate MBP threonine residue(s) and weakly phosphorylated MBP serine residue(s). Of the divalent cations tested, Mg(2+) was required for CfSAPK activation. Phosphatase treatment of CfSAPK abolished its kinase activity, indicating that phosphorylation is required for CfSAPK activation. Seven internal tryptic peptides identified from CfSAPK matched mitogen-activated protein kinases (MAPKs) in plants. CfSAPK cDNA was cloned using RT-PCR and rapid amplification of cDNA ends (RACE). CfSAPK cDNA encodes a 382-amino acid protein with a calculated molecular mass of 43,466.9 Da. The CfSAPK protein contains all 11 conserved kinase subdomains found in other Ser/Thr kinases. The amino acids sequence of CfSAPK is highly homologous to group A MAPKs in monocotyledon plants.
authors.affiliation: State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China.
authors.forename: Lubin
authors.initials: L
authors.lastname: Li
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.forename: Chunhui
authors.initials: C
authors.lastname: Xu
authors.forename: Tongbing
authors.initials: T
authors.lastname: Su
authors.forename: Dongtao
authors.initials: D
authors.lastname: Ren
authors.forename: Hailian
authors.initials: H
authors.lastname: Yang
mesh_terms: D031723:Bambusa / Q000201:enzymology*; D001483:Base Sequence; D017931:DNA Primers; D004789:Enzyme Activation; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D010766:Phosphorylation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
chemical_list: D017931:DNA Primers; D020928:Mitogen-Activated Protein Kinases