Washio K, Ishikawa K
Division of Biological Science, Graduate School of Environmental Earth Science, Hokkaido University, Sapporo, Japan.
Plant Physiol. 1994 Aug;105(4):1275-80. doi: 10.1104/pp.105.4.1275.
Several cDNA clones encoding either serine carboxypeptidases or related proteins of Oryza sativa L. were identified, and the abundance of the corresponding mRNA in immature and germinated grains was examined. The deduced amino acid sequence of each cDNA included key sequences, such as a pentapeptide (G-X-S-X-G/A) that is conserved among many serine carboxypeptidases, and the putative protein products were classified as two general and one novel type of cereal serine carboxypeptidases. Two general types exhibited considerable homology to type I and type III carboxypeptidases of cereal plants. The novel type encoded a serine carboxypeptidase-like protein that was very similar to type III carboxypeptidases of barley and wheat but had slight differences in both the N- and the C-terminal sequences. The mRNAs of each of these carboxypeptidases were observed in immature grains, and they decreased during maturation. The abundance of mRNA for each class of carboxypeptidase increased again following germination with the same time course and in a tissue-specific manner. The mRNAs for type I and type III-like carboxypeptidases were abundant in germinated embryos composed of leaf, root, and scutellum, whereas the mRNA for type III carboxypeptidase was conspicuous in endosperm that contained the aleurone layer. Altered amounts of mRNA in deembryonated half-grains in response to phytohormones, such as gibberellic acid and abscisic acid, were only detectable in the case of type III carboxypeptidase. Southern blot analysis using rice genomic DNA revealed the simple organization of each gene for these three classes of carboxypeptidases.
鉴定出了几个编码水稻丝氨酸羧肽酶或相关蛋白的cDNA克隆,并检测了未成熟和发芽谷粒中相应mRNA的丰度。每个cDNA推导的氨基酸序列包含关键序列,如许多丝氨酸羧肽酶中保守的五肽(G-X-S-X-G/A),推定的蛋白质产物被分类为两种常见类型和一种新型的谷物丝氨酸羧肽酶。两种常见类型与谷物植物的I型和III型羧肽酶表现出相当高的同源性。新型编码一种丝氨酸羧肽酶样蛋白,它与大麦和小麦的III型羧肽酶非常相似,但在N端和C端序列上都有细微差异。这些羧肽酶各自的mRNA在未成熟谷粒中都有观察到,并且在成熟过程中减少。随着发芽,每类羧肽酶的mRNA丰度再次增加,并具有相同的时间进程和组织特异性方式。I型和III型样羧肽酶的mRNA在由叶、根和盾片组成的发芽胚中丰富,但III型羧肽酶的mRNA在含有糊粉层的胚乳中很明显。仅在III型羧肽酶的情况下,才能检测到去胚半粒谷粒中响应植物激素(如赤霉素和脱落酸)而改变的mRNA量。使用水稻基因组DNA的Southern印迹分析揭示了这三类羧肽酶各自基因的简单结构。