Phean-O-Pas Srivilai, Punteeranurak Pornpimon, Buaboocha Teerapong
Department of Biochemistry, Faculty of Science, Chulalongkorn University, Payathai Road, Patumwan, Bangkok 10330, Thailand.
J Biochem Mol Biol. 2005 Jul 31;38(4):432-9. doi: 10.5483/bmbrep.2005.38.4.432.
Ca(2+)/calmodulin transduction pathways have been implicated in mediating stress response and tolerance in plants. Here, three genes encoding calmodulin (Cam) members of the EF-hand family of Ca(2+)-binding proteins were identified from Oryza sativa L. databases. Complementary DNA for each of the calmodulin genes, OsCam1, OsCam2, and OsCam3 were sequenced. OsCam1 and OsCam2 encode a conventional 148-amino acid calmodulin protein that contains four characteristic Ca(2+)-binding motifs. OsCam3 encode a similar protein with a 38-amino-acid extension containing a putative prenylation site (CVIL) at the carboxyl terminus. RT-PCR showed that each of the genes is expressed in leaves and roots of 2-week old rice seedlings. By RNA gel blot analysis, OsCam1 mRNA levels strongly increased in response to NaCl, mannitol and wounding treatments. In contrast, OsCam2 mRNA levels were relatively unchanged under all conditions investigated. NaCl treatment and wounding also increased the OsCam3 mRNA level, but in a more transient manner. Our results indicate that although the expression of genes encoding different calmodulin isoforms is ubiquitous, they are differentially regulated by various stress signals. In addition, we have demonstrated that the calcium-channel blocker lanthanum chloride inhibited the induction of OsCam1 gene expression by both NaCl and mannitol treatments. These results suggest that osmotic stressinduced expression of OsCam1 gene requires the [Ca(2+)]cyt elevation that is known to occur in response to these stimuli.
钙离子/钙调蛋白转导途径参与介导植物的应激反应和耐受性。在此,从水稻数据库中鉴定出三个编码钙离子结合蛋白EF-手型家族钙调蛋白(Cam)成员的基因。对每个钙调蛋白基因OsCam1、OsCam2和OsCam3的互补DNA进行了测序。OsCam1和OsCam2编码一种传统的148个氨基酸的钙调蛋白,含有四个特征性的钙离子结合基序。OsCam3编码一种类似的蛋白,在羧基末端有一个38个氨基酸的延伸,包含一个假定的异戊二烯化位点(CVIL)。RT-PCR显示,每个基因在2周龄水稻幼苗的叶片和根中均有表达。通过RNA凝胶印迹分析,OsCam1的mRNA水平在NaCl、甘露醇和创伤处理后显著增加。相比之下,在所有研究条件下,OsCam2的mRNA水平相对不变。NaCl处理和创伤也增加了OsCam3的mRNA水平,但更为短暂。我们的结果表明,尽管编码不同钙调蛋白异构体的基因表达普遍存在,但它们受到各种应激信号的差异调节。此外,我们还证明,钙通道阻滞剂氯化镧抑制了NaCl和甘露醇处理对OsCam1基因表达的诱导。这些结果表明,渗透胁迫诱导的OsCam1基因表达需要已知在这些刺激下发生的细胞质钙离子浓度升高。