Cloning, expression and N-terminal myristoylation of CpCPK1, a calcium-dependent protein kinase from zucchini (Cucurbita pepo L.)
Cloning, expression and N-terminal myristoylation of CpCPK1, a calcium-dependent protein kinase from zucchini (Cucurbita pepo L.)
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DOI：
10.1023/a:1006125918023
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发表时间：
1999-01-01
影响因子：
5.1
通讯作者：
Lomax, TL
中科院分区：
文献类型：
--
作者：
Ellard-Ivey, M;Hopkins, RB;Lomax, TL
我们从西葫芦(Cucurbita Pepo L.)中分离到一个编码钙依赖蛋白激酶(CDPK)基因的全长cDNA克隆(CpCDPK1)。预测的氨基酸序列与拟南芥CDPK基因家族成员有很高的相似性，特别是AtCPK1和AtCPK2。对CpCPKI在黄化和轻度生长的西葫芦幼苗中的稳态mRNA水平的Northern分析表明，该转录物在黄化的下胚轴中含量最丰富，总体表达受到光的抑制。与不同物种的CDPK基因家族其他成员的描述一样，CyCPK1克隆具有可能的N-末端肉豆蔻化序列。本研究利用定点突变和体外转录/翻译耦合系统证明了该基因编码的蛋白质是由植物N-肉豆蔻酰基转移酶特异性地进行肉豆蔻酰化的。这是一种CDPK蛋白的肉豆蔻酰化的首次证明，这可能有助于该蛋白定位于质膜的机制。
We have isolated a full-length cDNA clone (CpCDPK1) encoding a calcium-dependent protein kinase (CDPK) gene from zucchini (Cucurbita pepo L.). The predicted amino acid sequence of the cDNA shows a remarkably high degree of similarity to members of the CDPK gene family from Arabidopsis thaliana, especially AtCPK1 and AtCPK2.Northern analysis of steady-state mRNA levels for CpCPKI in etiolated and light-grown zucchini seedlings shows that the transcript is most abundant in etiolated hypocotyls and overall expression is suppressed by light.As described for other members of the CDPK gene family from different species, the CyCPK1 clone has a putative N-terminal myristoylation sequence. In this study, site-directed mutagenesis and an in vitro coupled transcription/translation system were used to demonstrate that the protein encoded by this cDNA is specifically myristoylated by a plant N-myristoyl transferase. This is the first demonstration of myristoylation of-a CDPK protein which may contribute to the mechanism by which this protein is localized to the plasma membrane.