Chen Y., Baum G., Fromm H.
Department of Plant Genetics, Weizmann Institute of Science, 76100 Rehovot, Israel.
Plant Physiol. 1994 Dec;106(4):1381-1387. doi: 10.1104/pp.106.4.1381.
A cDNA coding for a 58-kD calcium-dependent calmodulin (CaM)-binding glutamate decarboxylase (GAD) previously isolated in our laboratory from petunia (Petunia hybrida) (G. Baum, Y. Chen, T. Arazi, H. Takatsuji, H. Fromm [1993] J Biol Chem 268: 19610-19617) was used to conduct molecular studies of GAD expression. GAD expression was studied during petunia organ development using the GAD cDNA as a probe to detect the GAD mRNA and by the anti-recombinant GAD serum to monitor the levels of GAD. GAD activity was studied in extracts of organs in the course of development. The 58-kD CaM-binding GAD is expressed in all petunia organs tested (flowers and all floral parts, leaves, stems, roots, and seeds). The highest expression levels were in petals of open flowers. Developmental changes in the abundance of GAD mRNA and the 58-kD GAD were observed in flowers and leaves and during germination. Moreover, developmental changes in GAD activity in plant extracts coincided in most cases with changes in the abundance of the 58-kD GAD. We conclude that the 58-kD CaM-binding GAD is a ubiquitous protein in petunia organs and that its expression is developmentally regulated by transcriptional and/or posttranscriptional processes. Thus, GAD gene expression is likely to play a role in controlling the rates of GABA synthesis during petunia seed germination and organ development.
一个编码58-kD钙依赖性钙调蛋白(CaM)结合型谷氨酸脱羧酶(GAD)的cDNA,该cDNA先前在我们实验室从矮牵牛(Petunia hybrida)中分离得到(G. Baum、Y. Chen、T. Arazi、H. Takatsuji、H. Fromm [1993] J Biol Chem 268: 19610 - 19617),被用于进行GAD表达的分子研究。使用GAD cDNA作为探针检测GAD mRNA,并通过抗重组GAD血清监测GAD水平,从而在矮牵牛器官发育过程中研究GAD表达。在发育过程中,对器官提取物中的GAD活性进行了研究。58-kD CaM结合型GAD在所有测试的矮牵牛器官(花及所有花部、叶、茎、根和种子)中均有表达。最高表达水平出现在开放花的花瓣中。在花和叶以及种子萌发过程中,观察到了GAD mRNA丰度和58-kD GAD的发育变化。此外,植物提取物中GAD活性的发育变化在大多数情况下与58-kD GAD丰度的变化一致。我们得出结论,58-kD CaM结合型GAD是矮牵牛器官中一种普遍存在的蛋白质,其表达受转录和/或转录后过程的发育调控。因此,GAD基因表达可能在控制矮牵牛种子萌发和器官发育过程中γ-氨基丁酸(GABA)的合成速率方面发挥作用。