College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China.
J Integr Plant Biol. 2010 May;52(5):442-52. doi: 10.1111/j.1744-7909.2010.00906.x.
In search for components of mitogen-activated protein kinase (MAPK) cascades in maize (Zea mays) involved in response to abscisic acid (ABA) stimulus, a novel MAPK gene, ZmMPK3, from ABA-treated maize leaves cDNA was isolated and characterized. The full length of the ZmMPK3 gene is 1 520 bp and encodes a 376 amino acid protein with a predicted molecular mass of 43.5 kD and a pI of 5.83. ZmMPK3 contains all 11 MAPK conserved subdomains and the phosphorylation motif TEY. Amino acid sequence alignment revealed that ZmMPK3 shared high identity with group-A MAPK in plants. A time course (30-360 min) experiment using a variety of signal molecules and stresses revealed that the transcripts level of ZmMPK3 accumulated markedly and rapidly when maize seedlings were subjected to exogenous signaling molecules: ABA, H2O2, jasmonic acid and salicylic acid, various abiotic stimuli such as cold, drought, ultraviolet light, salinity, heavy metal and mechanical wounding. Its transcription was also found to be tissue-specific regulated. Here, we show that ABA and H2O2 induced a significant increase in the ZmMPK3 activity using immunoprecipitation and in-gel kinase assay. Furthermore, the results showed that the ZmMPK3 protein is localized mainly to the nucleus. These results suggest that the ZmMPK3 may play an important role in response to environmental stresses.
为了寻找玉米(Zea mays)中参与脱落酸(ABA)刺激反应的丝裂原活化蛋白激酶(MAPK)级联反应的成分,从ABA 处理的玉米叶片 cDNA 中分离并鉴定了一种新型的 MAPK 基因 ZmMPK3。ZmMPK3 基因的全长为 1520bp,编码一个 376 个氨基酸的蛋白质,预测分子量为 43.5kD,等电点为 5.83。ZmMPK3 含有所有 11 个 MAPK 保守亚结构域和磷酸化基序 TEY。氨基酸序列比对表明,ZmMPK3 与植物中的 A 组 MAPK 具有高度同源性。使用各种信号分子和应激物的时间过程(30-360min)实验表明,当玉米幼苗受到外源信号分子:ABA、H2O2、茉莉酸和水杨酸、各种非生物胁迫如寒冷、干旱、紫外线、盐度、重金属和机械损伤时,ZmMPK3 的转录水平明显且迅速积累。还发现其转录受到组织特异性调控。在这里,我们表明 ABA 和 H2O2 通过免疫沉淀和凝胶激酶测定诱导 ZmMPK3 活性显著增加。此外,结果表明 ZmMPK3 蛋白主要定位于细胞核。这些结果表明 ZmMPK3 可能在应对环境胁迫中发挥重要作用。