PNPase活性决定了叶绿体中mRNA 3'末端加工的效率、tRNA的降解以及多聚腺苷酸化的程度。
PNPase activity determines the efficiency of mRNA 3'-end processing, the degradation of tRNA and the extent of polyadenylation in chloroplasts.
作者信息
Walter Michael, Kilian Joachim, Kudla Jörg
机构信息
Molekulare Botanik, Universität Ulm, Albert-Einstein-Allee 11, D-89069 Ulm, Germany.
出版信息
EMBO J. 2002 Dec 16;21(24):6905-14. doi: 10.1093/emboj/cdf686.
Abstract
The exoribonuclease polynucleotide phosphorylase (PNPase) has been implicated in mRNA processing and degradation in bacteria as well as in chloroplasts of higher plants. Here, we report the first comprehensive in vivo study of chloroplast PNPase function. Modulation of PNPase activity in Arabidopsis chloroplasts by a reverse genetic approach revealed that, although this enzyme is essential for efficient 3'-end processing of mRNAs, it is insufficient to mediate transcript degradation. Surprisingly, we identified PNPase as also being indispensable for 3'-end maturation of 23S rRNA transcripts. Analysis of tRNA amounts in transgenic Arabidopsis plants suggests a direct correlation of PNPase activity and tRNA levels, indicating an additional function of this exoribo nuclease in tRNA decay. Moreover, the extent of polyadenylated mRNAs in chloroplasts is negatively correlated with PNPase activity. Together, our results attribute novel functions to PNPase in the metabolism of all major classes of plastid RNAs and suggest an unexpectedly complex role for PNPase in RNA processing and decay.
摘要
外切核糖核酸酶多核苷酸磷酸化酶（PNPase）已被证明参与细菌以及高等植物叶绿体中的mRNA加工和降解过程。在此，我们报道了第一项关于叶绿体PNPase功能的全面体内研究。通过反向遗传学方法调节拟南芥叶绿体中PNPase的活性表明，尽管这种酶对于mRNA的有效3'末端加工至关重要，但它不足以介导转录本的降解。令人惊讶的是，我们发现PNPase对于23S rRNA转录本的3'末端成熟也是不可或缺的。对转基因拟南芥植物中tRNA含量的分析表明，PNPase活性与tRNA水平直接相关，这表明这种外切核糖核酸酶在tRNA降解中具有额外功能。此外，叶绿体中多聚腺苷酸化mRNA的程度与PNPase活性呈负相关。总之，我们的结果赋予了PNPase在所有主要类别的质体RNA代谢中的新功能，并表明PNPase在RNA加工和降解中具有出人意料的复杂作用。
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