Auxin and heat shock activation of a novel member of the calmodulin like domain protein kinase gene family in cultured alfalfa cells
Auxin and heat shock activation of a novel member of the calmodulin like domain protein kinase gene family in cultured alfalfa cells
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DOI：
10.1093/jexbot/52.355.215
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发表时间：
2001-02-01
影响因子：
6.9
通讯作者：
Deák, M
中科院分区：
文献类型：
--
作者：
Davletova, S;Mészáros, T;Deák, M
在苜蓿中鉴定了钙调素样结构域蛋白激酶（CPK）同源物，并将其命名为MsCPK 3。cDNA全长序列编码一个535个氨基酸的多肽，分子量为60.2 kDa。推导的氨基酸序列显示了该激酶家族其他成员的所有保守基序，如丝氨酸-苏氨酸激酶结构域，一个连接区和四个潜在的Ca 2+结合EF位点。重组MsCPK 3蛋白从大肠杆菌中纯化。在体外磷酸化实验中，Ca ~（2+）可激活大肠杆菌的细胞，而钙调素拮抗剂（W-7）则可抑制这种作用。MsCPK 3基因在2，4-D诱导苜蓿体细胞胚胎发生的早期表达量增加。热激也激活了该基因，而激动素、阿坝和NaCl处理没有导致MsCPK 3 mRNA的积累。提出的数据表明，新的苜蓿CPK不同的压力反应，从以前描述的同系物，并在其潜在的参与激素和应力激活的重编程的发育途径在体细胞胚胎发生。
A calmodulin like domain protein kinase (CPK) homologue was identified in alfalfa and termed MsCPK3. The full-length sequence of cDNA encoded a 535 amino acid polypeptide with a molecular weight of 60.2 kDa. The deduced amino acid sequence showed all the conserved motifs that define other members of this kinase family, such as serine-threonine kinase domain, a junction region and four potential Ca2+-binding EF sites. The recombinant MsCPK3 protein purified from E. coli was activated by Ca2+ and inhibited by calmodulin antagonist (W-7) in vitro phosphorylation assays. The expression of MsCPK3 gene increased in the early phase of the 2,4-D induced alfalfa somatic embryogenesis. Heat shock also activated this gene while kinetin, ABA and NaCl treatment did not result in MsCPK3 mRNA accumulation. The data presented suggest that the new alfalfa CPK differs in stress responses from the previously described homologues and in its potential involvement in hormone and stress-activated reprogramming of developmental pathways during somatic embryogenesis.