Chan Chui Sien, Peng Hsiao-Ping, Shih Ming-Che
Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242, USA.
Plant Physiol. 2002 Nov;130(3):1476-86. doi: 10.1104/pp.007849.
Expression of nuclear genes that encode the A and B subunits of chloroplast glyceraldehyde-3-phosphate dehydrogenase (GAPA and GAPB) of Arabidopsis is known to be regulated by light. We used a negative selection approach to isolate mutants that were defective in light-regulated expression of the GAPA gene. Two dominant mutants belonging to the same complementation group, uga1-1 and uga1-2, were then characterized. These two mutants showed a dramatic reduction in GAPA mRNA level in both mature plants and seedlings. Surprisingly, mutations in uga1-1 and uga1-2 had no effect on the expression of GAPB and several other light-regulated genes. In addition, we found that the chloroplast glyceraldehyde-3-phosphate dehydrogenase enzyme activity of the mutants was only slightly lower than that of the wild type. Western-blot analysis showed that the GAPA protein level was nearly indistinguishable between the wild-type and the uga mutants. These results suggested that posttranscriptional control was involved in the up-regulation of the GAPA protein in the mutants. The uga1-1 mutation was mapped to the bottom arm of chromosome V of the Arabidopsis genome.
已知拟南芥叶绿体甘油醛-3-磷酸脱氢酶(GAPA和GAPB)的A和B亚基编码核基因的表达受光调控。我们采用负选择方法分离出在GAPA基因光调控表达方面存在缺陷的突变体。随后对属于同一互补群的两个显性突变体uga1-1和uga1-2进行了表征。这两个突变体在成熟植株和幼苗中GAPA mRNA水平均显著降低。令人惊讶的是,uga1-1和uga1-2中的突变对GAPB及其他几个光调控基因的表达没有影响。此外,我们发现突变体的叶绿体甘油醛-3-磷酸脱氢酶活性仅略低于野生型。蛋白质免疫印迹分析表明,野生型和uga突变体之间GAPA蛋白水平几乎没有差异。这些结果表明,转录后调控参与了突变体中GAPA蛋白的上调。uga1-1突变被定位到拟南芥基因组第五条染色体的短臂上。