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扩展鱼腥藻(念珠藻)PCC 7120 中的 FurC(PerR)调控基因簇:全基因组鉴定新的直接靶标揭示 FurC 参与中心碳代谢调控。

Expanding the FurC (PerR) regulon in Anabaena (Nostoc) sp. PCC 7120: Genome-wide identification of novel direct targets uncovers FurC participation in central carbon metabolism regulation.

机构信息

Departamento de Bioquímica y Biología Molecular y Celular and Institute for Biocomputation and Physics of Complex Systems, Universidad de Zaragoza, Zaragoza, Spain.

出版信息

PLoS One. 2023 Aug 7;18(8):e0289761. doi: 10.1371/journal.pone.0289761. eCollection 2023.

Abstract

FurC (PerR, Peroxide Response Regulator) from Anabaena sp. PCC 7120 (also known as Nostoc sp. PCC 7120) is a master regulator engaged in the modulation of relevant processes including the response to oxidative stress, photosynthesis and nitrogen fixation. Previous differential gene expression analysis of a furC-overexpressing strain (EB2770FurC) allowed the inference of a putative FurC DNA-binding consensus sequence. In the present work, more data concerning the regulon of the FurC protein were obtained through the searching of the putative FurC-box in the whole Anabaena sp. PCC 7120 genome. The total amount of novel FurC-DNA binding sites found in the promoter regions of genes with known function was validated by electrophoretic mobility shift assays (EMSA) identifying 22 new FurC targets. Some of these identified targets display relevant roles in nitrogen fixation (hetR and hgdC) and carbon assimilation processes (cmpR, glgP1 and opcA), suggesting that FurC could be an additional player for the harmonization of carbon and nitrogen metabolisms. Moreover, differential gene expression of a selection of newly identified FurC targets was measured by Real Time RT-PCR in the furC-overexpressing strain (EB2770FurC) comparing to Anabaena sp. PCC 7120 revealing that in most of these cases FurC could act as a transcriptional activator.

摘要

来自鱼腥藻 PCC 7120(也称为念珠藻 PCC 7120)的 FurC(PerR,过氧化物响应调节剂)是一种主要的调节剂,参与调节包括对氧化应激、光合作用和固氮的反应等相关过程。先前对 FurC 过表达菌株(EB2770FurC)的差异基因表达分析推断出了一个假定的 FurC DNA 结合保守序列。在本工作中,通过在鱼腥藻 PCC 7120 全基因组中搜索假定的 FurC-box,获得了更多关于 FurC 蛋白调控组的信息。通过电泳迁移率变动分析(EMSA)验证了在具有已知功能的基因启动子区域中发现的新型 FurC-DNA 结合位点的总量,鉴定了 22 个新的 FurC 靶标。这些鉴定的靶标中的一些在固氮(hetR 和 hgdC)和碳同化过程(cmpR、glgP1 和 opcA)中具有相关作用,表明 FurC 可能是协调碳氮代谢的另一个因素。此外,通过在 FurC 过表达菌株(EB2770FurC)中比较实时 RT-PCR 测量了一组新鉴定的 FurC 靶标的差异基因表达,结果表明在大多数情况下 FurC 可以作为转录激活剂。

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