Cloning and biochemical properties of CDPK gene OsCDPK14 from rice

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• 作者：Tiegang Zhang ; Qi Wang ; Xing Chen ; Chunhua Tian ; Xinshen Wang ; Tim Xing ; Yi Li ; Yingdian Wang
• 关键词：Calcium-dependent protein kinase (CDPK) ; cDNA cloning ; Kinase activities ; OsCDPK14 ; Rice (Oryza sativa) ; Subcellular localization
• 刊名：Journal of Plant Physiology
• 出版年：2005
• 出版时间：14 October 2005
• 年：2005
• 卷：162
• 期：10
• 页码：1149-1159
• 全文大小：754 K

文摘

A rice CDPK gene, OsCDPK14 (AY144497), was cloned from developing caryopses of rice (Oryza sativa cv. Zhonghua 15). Its cDNA sequence (1922 bp) contains an ORF encoding a 514 amino acids protein (56.7 kD, pl 5.18). OsCDPK14 shows the typical structural features of the CDPK family, including a conserved catalytic Ser/Thr kinase domain, an autoinhibitory domain and a CaM-like domain with four putative Ca²⁺-binding EF hands. Subcellular targeting indicated that OsCDPK14 was located in the cytoplasm, probably due to the absence of myristoylation and palmitoylation motifs. OsCDPK14 was expressed in Escherichia coli and purified from bacterial extracts. The recombinant protein was shown to be a functional protein kinase using Syntide-2, a synthetic peptide. Kinase activity was shown to be Ca²⁺-dependent, and this activation was strongly enhanced by Mn²⁺ and inhibited by W7 in vitro. These results provide significant insights into the regulation and biochemical properties of OsCDPK14, suggesting OsCDPK14 may be a signal factor of cytoplasm in rice plant.