Woodger Fiona J, Gubler Frank, Pogson Barry J, Jacobsen John V
CSIRO Plant Industry, GPO Box 1600, Canberra, ACT, 2601, Australia.
Plant J. 2003 Feb;33(4):707-17. doi: 10.1046/j.1365-313x.2003.01663.x.
GAMYB is a gibberellin (GA)-regulated activator of hydrolase gene expression in the aleurone layer of germinating cereal grains. Although it is clear that GAMYB expression is regulated by GA, more remains to be understood about how this transcription factor operates within the GA-response pathway. In order to isolate new components from the GA-response pathway, barley aleurone libraries were screened for GAMYB-binding proteins using a recently developed yeast two-hybrid system, which is compatible with the use of transcription factors as baits. We isolated a new member of the emerging Mak-subgroup of cdc2- and MAP kinase-related protein kinases. We have termed this GAMYB-binding protein KGM (for kinase associated with GAMYB). Transient expression of KGM specifically repressed alpha-amylase promoter activity at the level of GAMYB function but a mutation designed to de-stabilise the activation loop of KGM alleviated this repression. We propose that KGM is a negative regulator of GAMYB function in aleurone that may prevent precocious hydrolase gene expression.
GAMYB是一种受赤霉素(GA)调控的水解酶基因表达激活因子,存在于发芽谷物的糊粉层中。尽管GAMYB的表达受GA调控这一点很明确,但关于该转录因子在GA反应途径中如何发挥作用,仍有许多有待了解之处。为了从GA反应途径中分离出新的组分,利用最近开发的酵母双杂交系统筛选大麦糊粉层文库中与GAMYB结合的蛋白,该系统适用于以转录因子作为诱饵。我们分离出了一个新的与细胞周期蛋白依赖性激酶2(cdc2)和丝裂原活化蛋白激酶(MAPK)相关的蛋白激酶家族Mak亚组成员。我们将这个与GAMYB结合的蛋白命名为KGM(与GAMYB相关的激酶)。KGM的瞬时表达在GAMYB功能水平上特异性抑制了α-淀粉酶启动子活性,但设计用于使KGM激活环不稳定的突变减轻了这种抑制作用。我们提出,KGM是糊粉层中GAMYB功能的负调节因子,可能会阻止水解酶基因的过早表达。