水稻中水杨酸和茉莉酸特异诱导蛋白激酶基因OsSJMK1的分离和鉴定(英文)

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裂原激活蛋白激酶(MAPK)级联系统负责把接受自胞外或胞内的信号进一步传递和放大而最终作用于特异的转录因子,从而启动或调控基因的表达。MAPK信号级联系统在细胞分裂、分化、生物胁迫和非生物胁迫等多种信号传递途径中起着十分重要的作用。该文以一个茉莉酸(JA)诱导的表达序列为基础,在水稻中分离到了一个裂原激活蛋白激酶基因OsSJMK1的全长cDNA。序列比较分析表明该基因编码498个氨基酸,蛋白质等电点为8.43,包括完整的MAPK家族的蛋白激酶结构域。OsSJMK1与所有物种的MAPK一样包括蛋白激酶的全部11个次级结构域,在VII和VIII次级结构域间一个双磷酸化位点;该位点苏氨酸(T)和酪氨酸(Y)残基之间为天冬氨酸(D),而不是其他MAPK中常见的谷氨酸(E)、脯氨酸(P)或甘氨酸(G)。除了典型的MAPK激酶功能域外,在羧基端还有一段长约150个氨基酸残基的可能参与蛋白互作的结构域。以上这些结构特征表明OsSJMK1属于植物中第V类MAPK家族成员。蛋白激酶结构域序列比较表明OsSJMK1与报道的稻瘟病菌和机械伤害诱导的BWMK1的序列相似性高达81%,而且基因内含子和外显子的组成也非常相似,属于同一亚类,但在蛋白质序列的C端差异却很大。与BWMK1不同,OsSJMK1的表达不受伤害诱导,而受稻瘟病轻微诱导,但在JA和SA(水杨酸)处理早期表达量却迅速升高。在JA处理后1h,OsSJMK1转录水平升高到最大,而12h后回落到处理前的本底水平;在SA处理后30min转录水平就开始上升,2h达到最高值,而随后开始下降到处理前的本底水平。SA类似物BTH也能诱导OsSJMK1的表达。其他一些激素处理(如ABA)和非生物胁迫(如干旱、盐胁迫)都不对基因的表达产生任何影响,而且在植物大部分组织中的表达量都非常低。这些结果说明OsSJMK1可能特异性的参与JA和SA介导的防卫反应。 The mitogen-activated protein kinase (MAPK) cascade system is responsible for initiating or regulating the expression of genes by further transmitting and amplifying signals that are either taken from outside or inside the cell and ultimately act on specific transcription factors. MAPK signal cascade system plays an important role in many signal transduction pathways such as cell division, differentiation, biotic stress and abiotic stress. Based on a jasmonic acid (JA) -induced expression sequence, a full-length cDNA of OsSJMK1, a mitogen-activated protein kinase gene, was isolated from rice. Sequence comparison analysis showed that the gene encoded 498 amino acids, the protein isoelectric point was 8.43, including the complete MAPK family protein kinase domain. OsSJMK1, like MAPKs of all species, includes all 11 secondary domains of protein kinases and a double phosphorylation site between the VII and VIII secondary domains; this site contains threonine (T) and tyrosine ) Residue is aspartic acid (D) rather than glutamic acid (E), proline (P) or glycine (G) as is common in other MAPKs. In addition to the typical MAPK kinase domain, a further 150 amino acid residue at the carboxy terminus is involved in the protein-protein interaction domain. These structural features indicate that OsSJMK1 belongs to the class V MAPK family member in plants. Comparison of protein kinase domain sequences showed that the sequence similarity of OsSJMK1 to reported M. oryzae and BWMK1 induced by mechanical injury was as high as 81%, and the composition of introns and exons was also very similar, belonging to the same subgroup, but at The C-terminal differences in protein sequences are substantial. Unlike BWMK1, OsSJMK1 expression was not induced by injury but slightly induced by rice blast but rapidly increased in the early stage of JA and SA (salicylic acid) treatment. At 1 h after JA treatment, the transcription level of OsSJMK1 increased to the maximum, and returned to the baseline level after 12 h. The transcription level began to rise 30 min after SA treatment and reached the peak at 2 h, and then decreased to the level before treatment The background level. The SA analogue BTH also induced OsSJMK1 expression. Some other hormonal treatments (such as ABA) and abiotic stresses (such as drought and salt stress) have no effect on gene expression, and are expressed very low in most plant tissues. These results indicate that OsSJMK1 may be specifically involved in JA and SA-mediated defense responses.
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