The Hfq regulon of Neisseria meningitidis
The Hfq regulon of Neisseria meningitidis
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DOI：
10.1002/2211-5463.12218
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发表时间：
2017-06-01
期刊：
影响因子：
2.6
通讯作者：
Pannekoek, Yvonne
中科院分区：
文献类型：
--
作者：
in't Veld, Robert A. G. Huis;Kramer, Gertjan;Pannekoek, Yvonne
保守的RNA结合蛋白Hfq在原核细胞内具有多种调节作用，包括促进小RNA和mRNA之间稳定的双链体形成，因此Hfq缺失突变体具有多效性表型。以前的脑膜炎奈瑟菌蛋白质组和转录组研究产生有限的洞察差异基因表达由于Hfq的损失。在这项研究中，反相液相色谱结合数据独立的交替扫描质谱（LC-MSE）用于快速高分辨率定量蛋白质组学分析，以进一步阐明脑膜炎球菌hfq缺失突变体的差异表达蛋白质组。N.对在液体生长培养基中生长的脑膜炎血清群B H44/76野生型（wt）和H44/76 Δ hfq（Δ hfq）进行胰蛋白酶消化。在通过质谱法（MSE）分析之前，通过液相色谱法（LC）分离所得肽混合物。通过学生t检验分析差异表达，并以假发现率（FDR）为对照。用506种蛋白质（20%）实现了比较wt和Δ hfq的相对表达的可靠定量。FDR控制时，q
The conserved RNA-binding protein, Hfq, has multiple regulatory roles within the prokaryotic cell, including promoting stable duplex formation between small RNAs and mRNAs, and thus hfq deletion mutants have pleiotropic phenotypes. Previous proteome and transcriptome studies of Neisseria meningitidis have generated limited insight into differential gene expression due to Hfq loss. In this study, reversed-phase liquid chromatography combined with data-independent alternate scanning mass spectrometry (LC-MSE) was utilized for rapid high-resolution quantitative proteomic analysis to further elucidate the differentially expressed proteome of a meningococcal hfq deletion mutant. Whole-cell lysates of N. meningitidis serogroup B H44/76 wild-type (wt) and H44/76 Delta hfq (Delta hfq) grown in liquid growth medium were subjected to tryptic digestion. The resulting peptide mixtures were separated by liquid chromatography (LC) prior to analysis by mass spectrometry (MSE). Differential expression was analyzed by Student's t-test with control for false discovery rate (FDR). Reliable quantitation of relative expression comparing wt and Delta hfq was achieved with 506 proteins (20%). Upon FDR control at q