Functional characterization of the principal sigma factor RpoD of phytoplasmas via an in vitro transcription assay.
Functional characterization of the principal sigma factor RpoD of phytoplasmas via an in vitro transcription assay.
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DOI：
10.1038/srep11893
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发表时间：
2015-07-07
影响因子：
4.6
通讯作者：
Namba S
中科院分区：
文献类型：
--
作者：
Miura C;Komatsu K;Maejima K;Nijo T;Kitazawa Y;Tomomitsu T;Yusa A;Himeno M;Oshima K;Namba S
植物原体（Mollicutes纲）是一种昆虫传播和植物致病菌，通过寄主转换在植物和昆虫的细胞内繁殖。我们前期的研究发现，y - m菌株的植物浆sigma因子rpoD （rpoDOY）可能是调控寄主转换的关键因子，因为rpoDOY在昆虫寄主中的表达水平高于植物寄主。在这项研究中，我们开发了一种体外转录测定系统来鉴定rpodoy依赖基因和共识启动子元件。结果表明，RpoDOY调控了y - m菌株的一些内源性、毒力和宿主-植物原体相互作用基因。这些基因转录起始位点上游区域包含保守的-35和-10启动子序列，与典型的细菌rpod依赖启动子元件相似，而-35启动子元件是可变的。此外，我们利用计算机工具在植物原体全基因组序列上搜索基于这些启动子元件的推测的rpod依赖基因。植物原体RpoD似乎不仅介导了许多作为主要sigma因子的管家基因的转录，而且还介导了具有宿主特异性表达模式的毒力和宿主-植物原体相互作用相关基因的转录。这些结果表明，在使植物原体转换宿主的基因调控方面，存在比以前认为的更复杂的机制。
Phytoplasmas (class, Mollicutes) are insect-transmissible and plant-pathogenic bacteria that multiply intracellularly in both plants and insects through host switching. Our previous study revealed that phytoplasmal sigma factor rpoD of OY-M strain (rpoDOY) could be a key regulator of host switching, because the expression level of rpoDOY was higher in insect hosts than in plant hosts. In this study, we developed an in vitro transcription assay system to identify RpoDOY-dependent genes and the consensus promoter elements. The assay revealed that RpoDOY regulated some housekeeping, virulence, and host–phytoplasma interaction genes of OY-M strain. The upstream region of the transcription start sites of these genes contained conserved –35 and –10 promoter sequences, which were similar to the typical bacterial RpoD-dependent promoter elements, while the –35 promoter elements were variable. In addition, we searched putative RpoD-dependent genes based on these promoter elements on the whole genome sequence of phytoplasmas using in silico tools. The phytoplasmal RpoD seems to mediate the transcription of not only many housekeeping genes as the principal sigma factor, but also the virulence- and host-phytoplasma interaction-related genes exhibiting host-specific expression patterns. These results indicate that more complex mechanisms exist than previously thought regarding gene regulation enabling phytoplasmas to switch hosts.
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影响因子：
4.4
作者：
Andersen MT;Liefting LW;Havukkala I;Beever RE
通讯作者：
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作者：
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通讯作者：
NOMURA, M