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烟草质体中rbcL转录本水平与光照无关:黑暗中转录速率降低可通过mRNA稳定性增加得到补偿。

rbcL Transcript levels in tobacco plastids are independent of light: reduced dark transcription rate is compensated by increased mRNA stability.

作者信息

Shiina T, Allison L, Maliga P

机构信息

Waksman Institute, Rutgers, The State University of New Jersey, 190 Frelinghuysen Road, Piscataway, New Jersey 08854-8020, USA.

出版信息

Plant Cell. 1998 Oct;10(10):1713-22. doi: 10.1105/tpc.10.10.1713.

Abstract

The plastid rbcL gene, encoding the large subunit of ribulose-1, 5-bisphosphate carboxylase, in higher plants is transcribed from a sigma70 promoter by the eubacterial-type RNA polymerase. To identify regulatory elements outside of the rbcL -10/-35 promoter core, we constructed transplastomic tobacco plants with uidA reporter genes expressed from rbcL promoter derivatives. Promoter activity was characterized by measuring steady state levels of uidA mRNA on RNA gel blots and by measuring promoter strength in run-on transcription assays. We report here that the rbcL core promoter is sufficient to obtain wild-type rates of transcription. Furthermore, the rates of transcription were up to 10-fold higher in light-grown leaves than in dark-adapted plants. Although the rates of transcription were lower in the dark, rbcL mRNA accumulated to similar levels in light-grown and dark-adapted leaves. Accumulation of uidA mRNA from most rbcL promoter deletion derivatives directly reflected the relative rates of transcription: high in the light-grown and low in the dark-adapted leaves. However, uidA mRNA accumulated to high levels in a light-independent fashion as long as a segment encoding a stem-loop structure in the 5' untranslated region was included in the promoter construct. This finding indicates that lower rates of rbcL transcription in the dark are compensated by increased mRNA stability.

摘要

高等植物中编码1,5-二磷酸核酮糖羧化酶大亚基的质体rbcL基因,由真细菌型RNA聚合酶从sigma70启动子转录。为了鉴定rbcL -10/-35启动子核心区域以外的调控元件,我们构建了用rbcL启动子衍生物表达uidA报告基因的转基因烟草植株。通过在RNA凝胶印迹上测量uidA mRNA的稳态水平以及在连续转录分析中测量启动子强度来表征启动子活性。我们在此报告,rbcL核心启动子足以获得野生型转录速率。此外,光照下生长的叶片中的转录速率比暗适应植物中的高10倍。尽管黑暗中的转录速率较低,但rbcL mRNA在光照下生长和暗适应的叶片中积累到相似水平。大多数rbcL启动子缺失衍生物的uidA mRNA积累直接反映了相对转录速率:光照下生长的叶片中高,暗适应的叶片中低。然而,只要启动子构建体中包含5'非翻译区中编码茎环结构的片段,uidA mRNA就以不依赖光照的方式积累到高水平。这一发现表明,黑暗中rbcL转录速率较低可通过增加mRNA稳定性来补偿。

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