Analysis of a photosynthetic cyanobacterium rich in internal membrane systems via gradient profiling by sequencing (Grad-seq).
Analysis of a photosynthetic cyanobacterium rich in internal membrane systems via gradient profiling by sequencing (Grad-seq).
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通过测序梯度分析（Grad-seq）分析富含内膜系统的光合蓝藻。
DOI：
10.1093/plcell/koaa017
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发表时间：
2021-04-17
期刊：
影响因子：
--
通讯作者：
Hess WR
中科院分区：
文献类型：
--
作者：
Riediger M;Spät P;Bilger R;Voigt K;Maček B;Hess WR
尽管在光合作用的蓝藻中已经报道了调节小RNA，但缺乏明确的RNA伴侣参与其调节构成了一个难题。在这里，我们通过测序（Grad-seq）梯度分析了Synechocystis 6803的细胞rna和蛋白质的全部补体。可以区分具有重叠亚基的复合物，如cpcg1型与cpcl型藻胆体或PsaK1与PsaK2光系统I前（复合物），支持该方法的高质量。在梯度分布概况的聚类之后，通过几个额外的标准产生了潜在的RNA伴侣的短列表，包括YlxR同源物和KhpA/B复合物的蓝藻同源物。这些数据表明，以前未检测到的辅助蛋白和CRISPR-Cas系统之间的复合物，如Csx1-Csm6核糖核溶解防御复合物。此外，RpoZ或6S RNA与核心RNA聚合酶复合物单独结合，并且存在非活性sigma - anti - sigma复合物库。Synechocystis gradseq资源可在https://sunshine.biologie.uni-freiburg.de/GradSeqExplorer/上在线获得，为光合生物中rna -蛋白质复合物和多亚基蛋白质复合物的功能分配提供了全面的资源。我们使用Grad-seq分析了一种蓝藻，为深入分析光合生物中的复杂体提供了全面的资源。
Although regulatory small RNAs have been reported in photosynthetic cyanobacteria, the lack of clear RNA chaperones involved in their regulation poses a conundrum. Here, we analyzed the full complement of cellular RNAs and proteins using gradient profiling by sequencing (Grad-seq) in Synechocystis 6803. Complexes with overlapping subunits such as the CpcG1-type versus the CpcL-type phycobilisomes or the PsaK1 versus PsaK2 photosystem I pre(complexes) could be distinguished, supporting the high quality of this approach. Clustering of the in-gradient distribution profiles followed by several additional criteria yielded a short list of potential RNA chaperones that include an YlxR homolog and a cyanobacterial homolog of the KhpA/B complex. The data suggest previously undetected complexes between accessory proteins and CRISPR-Cas systems, such as a Csx1-Csm6 ribonucleolytic defense complex. Moreover, the exclusive association of either RpoZ or 6S RNA with the core RNA polymerase complex and the existence of a reservoir of inactive sigma–antisigma complexes is suggested. The Synechocystis Grad-seq resource is available online at https://sunshine.biologie.uni-freiburg.de/GradSeqExplorer/ providing a comprehensive resource for the functional assignment of RNA–protein complexes and multisubunit protein complexes in a photosynthetic organism. We analyze a cyanobacterium using Grad-seq, providing a comprehensive resource for the in-depth analysis of the complexome in a photosynthetic organism.
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影响因子：
3.3
作者：
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通讯作者：
Pieper, Jorg
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Georg, Jens;Dienst, Dennis;Wilde, Annegret
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5
作者：
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通讯作者：
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