Khandal Dhriti, Samol Iga, Buhr Frank, Pollmann Stephan, Schmidt Holger, Clemens Stephan, Reinbothe Steffen, Reinbothe Christiane
Université Joseph Fourier, CERMO, PO Box 53, F-38041 Grenoble Cedex 9, France.
Proc Natl Acad Sci U S A. 2009 Aug 4;106(31):13112-7. doi: 10.1073/pnas.0903522106. Epub 2009 Jul 20.
The tigrina (tig)-d.12 mutant of barley is impaired in the negative control limiting excess protochlorophyllide (Pchlide) accumulation in the dark. Upon illumination, Pchlide operates as photosensitizer and triggers singlet oxygen production and cell death. Here, we show that both Pchlide and singlet oxygen operate as signals that control gene expression and metabolite accumulation in tig-d.12 plants. In vivo labeling, Northern blotting, polysome profiling, and protein gel blot analyses revealed a selective suppression of synthesis of the small and large subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase (RBCSs and RBCLs), the major light-harvesting chlorophyll a/b-binding protein of photosystem II (LHCB2), as well as other chlorophyll-binding proteins, in response to singlet oxygen. In part, these effects were caused by an arrest in translation initiation of photosynthetic transcripts at 80S cytoplasmic ribosomes. The observed changes in translation correlated with a decline in the phosphorylation level of ribosomal protein S6. At later stages, ribosome dissociation occurred. Together, our results identify translation as a major target of singlet oxygen-dependent growth control and cell death in higher plants.
大麦的tigrina (tig)-d.12突变体在黑暗中限制过量原叶绿素酸酯(Pchlide)积累的负调控方面存在缺陷。光照后,Pchlide作为光敏剂起作用,引发单线态氧的产生和细胞死亡。在此,我们表明Pchlide和单线态氧均作为信号,控制tig-d.12植物中的基因表达和代谢物积累。体内标记、Northern印迹、多核糖体分析和蛋白质凝胶印迹分析显示,响应单线态氧,1,5-二磷酸核酮糖羧化酶/加氧酶(RBCSs和RBCLs)的小亚基和大亚基、光系统II的主要捕光叶绿素a/b结合蛋白(LHCB2)以及其他叶绿素结合蛋白的合成受到选择性抑制。部分这些效应是由光合转录本在80S细胞质核糖体上的翻译起始停滞引起的。观察到的翻译变化与核糖体蛋白S6的磷酸化水平下降相关。在后期,核糖体发生解离。总之,我们的结果确定翻译是高等植物中单线态氧依赖性生长控制和细胞死亡的主要靶点。