Initiating nucleotide identity determines efficiency of RNA synthesis from 6S RNA templates in Bacillus subtilis but not Escherichia coli.

Initiating nucleotide identity determines efficiency of RNA synthesis from 6S RNA templates in Bacillus subtilis but not Escherichia coli.
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DOI:
10.1093/nar/gkt517
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发表时间:
2013-08
影响因子:
14.9
通讯作者:
Wassarman KM
Wassarman KM
中科院分区:
生物学2区
文献类型:
--
作者:
Cabrera-Ostertag IJ;Cavanagh AT;Wassarman KM

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6S核糖核酸是一种非编码的小核糖核酸,它结合在管家形式的核糖核酸聚合酶的活性部位(例如,在大肠杆菌中的Eσ70,在枯草杆菌中的EσA),并调节转录。在6S RNA从静止相向外生长的过程中,RNA聚合酶的有效释放依赖于以6S RNA为模板生成产物RNA(PRNA)。有趣的是,枯草杆菌有两个6S RNA,6S-1和6S-2,但在生长过程中,似乎只有6S-1 RNA被有效地用作合成PRNA的模板。在这里,我们证明了起始核苷酸的同一性对于枯草杆菌RNA聚合酶使用RNA模板特别重要。具体地说,三磷酸鸟苷(GTP)是有效合成PRNA所必需的,这从机制上解释了为什么6S-2 RNA不支持强劲的PRNA合成,因为它是由三磷酸腺苷(ATP)引发的。耐人寻味的是,E.ColiRNA聚合酶对启动核苷酸同一性没有强烈的偏好。这些观察结果突显了枯草杆菌和大肠杆菌RNA聚合酶在生化性质上的重要差异,特别是在有效利用RNA模板的能力方面,这也可能反映了这两种生物在GTP和ATP代谢方面的差异。
The 6S RNA is a non-coding small RNA that binds within the active site of housekeeping forms of RNA polymerases (e.g. Eσ70 in Escherichia coli, EσA in Bacillus subtilis) and regulates transcription. Efficient release of RNA polymerase from 6S RNA regulation during outgrowth from stationary phase is dependent on use of 6S RNA as a template to generate a product RNA (pRNA). Interestingly, B. subtilis has two 6S RNAs, 6S-1 and 6S-2, but only 6S-1 RNA appears to be used efficiently as a template for pRNA synthesis during outgrowth. Here, we demonstrate that the identity of the initiating nucleotide is particularly important for the B. subtilis RNA polymerase to use RNA templates. Specifically, initiation with guanosine triphosphate (GTP) is required for efficient pRNA synthesis, providing mechanistic insight into why 6S-2 RNA does not support robust pRNA synthesis as it initiates with adenosine triphosphate (ATP). Intriguingly, E. coli RNA polymerase does not have a strong preference for initiating nucleotide identity. These observations highlight an important difference in biochemical properties of B. subtilis and E. coli RNA polymerases, specifically in their ability to use RNA templates efficiently, which also may reflect the differences in GTP and ATP metabolism in these two organisms.
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