STRUCTURE AND EXPRESSION OF 2 GENES THAT ENCODE DISTINCT DROUGHT-INDUCIBLE CYSTEINE PROTEINASES IN ARABIDOPSIS-THALIANA
STRUCTURE AND EXPRESSION OF 2 GENES THAT ENCODE DISTINCT DROUGHT-INDUCIBLE CYSTEINE PROTEINASES IN ARABIDOPSIS-THALIANA
复制标题
DOI：
10.1016/0378-1119(93)90266-6
复制
发表时间：
1993-07-30
期刊：
影响因子：
3.5
通讯作者：
SHINOZAKI, K
中科院分区：
文献类型：
--
作者：
KOIZUMI, M;YAMAGUCHISHINOZAKI, K;SHINOZAKI, K
在拟南芥（Arabidopsis thaliana）脱水应答基因对应的9个cDNA克隆（RD）中，进一步分析了RD19和RD21两个克隆。Northern blot分析显示，RD19和RD21 mrna均未被脱落酸诱导。RD19和RD21 mRNA的合成均不受冷应激和热应激的影响。另一方面，在高盐条件下，RD19和RD21 mrna的转录都被强烈诱导，这表明RD19和RD21对应的基因可能是通过植物细胞渗透电位的变化而被诱导的。由两种RD cdna编码的推测蛋白RD19和RD21具有半胱氨酸蛋白酶（CysP）催化位点的典型氨基酸（aa）序列。RD21和RD19似乎含有在蛋白质分泌中起作用的信号肽。RD21含有一个类似于c端延伸肽的aa序列。系统进化树分析表明，推测的RD21和RD19蛋白是完全不同类型的CysP。基因组南方分析显示，每个基因家族至少包含两个不交叉杂交的成员。克隆了RD19和RD21对应的两个基因（分别为rd19A和rd21A），结构分析显示它们分别含有2个和4个内含子。内含子的数量和位置在基因之间存在差异，这支持了我们的假设，即rd19A和rd21A属于编码CysP的不同基因亚家族。通过引物延伸确定转录起始点。在这两个基因的启动子区域发现了两个保守序列。
Among nine cDNA clones (named RD) corresponding to genes that are responsive to dehydration in Arabidopsis thaliana, two clones, RD19 and RD21, were analyzed further. Northern blot analysis revealed that both the RD19 and RD21 mRNAs were not induced by abscisic acid. Neither RD19 nor RD21 mRNA synthesis was responsive to cold or to heat stress. On the other hand, transcription of both the RD19 and RD21 mRNAs was strongly induced under high-salt conditions, which suggests that the genes corresponding to RD19 and RD21 may be induced by changes in the osmotic potential of plant cells. Putative proteins, RD19 and RD21, encoded by two of the RD cDNAs have amino acid (aa) sequences typical of the catalytic sites of cysteine proteinases (CysP). RD21 and RD19 appeared to contain signal peptides that function in protein secretion. RD21 contains an aa sequence similar to that of the C-terminal extension peptide. Phylogenetic tree analysis indicated that the putative RD21 and RD19 proteins are quite different types of CysP. Genomic Southern analysis revealed that each gene family contains at least two members, which do not cross-hybridize. The two genes corresponding to RD19 and RD21 (rd19A and rd21A, respectively) were cloned and their structural analysis revealed the presence of two and four introns, respectively. The numbers and sites of introns differ between the genes, supporting our hypothesis that rd19A and rd21A belong to different subfamilies of genes encoding CysP. The transcription start points were determined by primer extension. Two conserved sequences were found in the promoter regions of the two genes.