Hansen E R, Petracek M E, Dickey L F, Thompson W F
Department of Botany, North Carolina State University, Raleigh, North Carolina 27695, USA.
Plant Physiol. 2001 Feb;125(2):770-8. doi: 10.1104/pp.125.2.770.
Ferredoxin-1 (Fed-1) mRNA contains an internal light response element (iLRE) that destabilizes mRNA when light-grown plants are placed in darkness. mRNAs containing this element dissociate from polyribosomes in the leaves of transgenic tobacco (Nicotiana tabacum) plants transferred to the dark for 2 d. Here, we report in vivo labeling experiments with a chloramphenicol acetyl transferase mRNA fused to the Fed-1 iLRE. Our data indicate that the Fed-1 iLRE mediates a rapid decline in translational efficiency and that iLRE-containing mRNAs dissociate from polyribosomes within 20 min after plants are transferred to darkness. Both events occur before the decline in mRNA abundance, and polyribosome association is rapidly reversible if plants are re-illuminated. These observations support a model in which Fed-1 mRNA in illuminated leaves is stabilized by its association with polyribosomes, and/or by translation. In darkness a large portion of the mRNA dissociates from polyribosomes and is subsequently degraded. We also show that a significant portion of total tobacco leaf mRNA is shifted from polyribosomal to non-polyribosomal fractions after 20 min in the dark, indicating that translation of other mRNAs is also rapidly down-regulated in response to darkness. This class includes some, but not all, cytoplasmic mRNAs encoding proteins involved in photosynthesis.
铁氧化还原蛋白-1(Fed-1)mRNA含有一个内部光反应元件(iLRE),当光照生长的植物置于黑暗中时,该元件会使mRNA不稳定。含有此元件的mRNA在转移至黑暗2天的转基因烟草(Nicotiana tabacum)植株叶片中会从多核糖体上解离。在此,我们报道了用与Fed-1 iLRE融合的氯霉素乙酰转移酶mRNA进行的体内标记实验。我们的数据表明,Fed-1 iLRE介导翻译效率迅速下降,并且含有iLRE的mRNA在植物转移至黑暗后20分钟内从多核糖体上解离。这两个事件都发生在mRNA丰度下降之前,并且如果植物重新光照,多核糖体结合是迅速可逆的。这些观察结果支持一种模型,即光照叶片中的Fed-1 mRNA通过与多核糖体结合和/或翻译而稳定。在黑暗中,大部分mRNA从多核糖体上解离,随后被降解。我们还表明,黑暗处理20分钟后,烟草叶片总mRNA的很大一部分从多核糖体部分转移到非多核糖体部分,这表明其他mRNA的翻译也会因黑暗而迅速下调。这一类包括一些但不是所有编码参与光合作用蛋白质的细胞质mRNA。