Global systems-level analysis of Hfq and SmpB deletion mutants in Salmonella: implications for virulence and global protein translation.
Global systems-level analysis of Hfq and SmpB deletion mutants in Salmonella: implications for virulence and global protein translation.
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DOI：
10.1371/journal.pone.0004809
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Smith RD
中科院分区：
文献类型：
--
作者：
Ansong C;Yoon H;Porwollik S;Mottaz-Brewer H;Petritis BO;Jaitly N;Adkins JN;McClelland M;Heffron F;Smith RD
利用样本匹配的转录组学和蛋白质组学测量，现在可以开始了解肠道细菌转录后调控程序的影响。在细菌中，转录后调控是由相对较少的RNA结合蛋白因子介导的，包括CSRA、Hfq和SmpB。沙门氏菌中这三个基因CSRA、HfQ和smpB中的任何一个基因的突变都会减弱小鼠的毒力，并无法在巨噬细胞中生存。CSRA具有明确定义的特异性，其基础是结合特定的mRNA序列来抑制翻译。然而，Hfq和SmpB调控的蛋白质并不是很清楚。以前的工作是通过对结合的RNA进行直接测序来纯化RNA-蛋白质复合体，并发现它与数量惊人的转录本结合，从而确定了受HfQ调控的蛋白质。在这份报告中，我们使用全球蛋白质组学通过比较亲本和同基因Hfq或SmpB突变体中的蛋白丰度来直接鉴定受Hfq或SmpB调控的蛋白质。我们还从这些样本中制备了用于微阵列分析的RNA，以确定蛋白质表达的变化是否是转录后介导的。对在四种不同条件下生长的细菌样本进行了分析；两种实验室条件和两种被认为模拟细胞内环境的条件。我们发现，hfq和smpB的突变体分别直接或间接调控了至少20%和4%的沙门氏菌蛋白，转录和蛋白表达之间的相关性有限。这些蛋白代表了许多生物学过程所需的广泛的沙门氏菌蛋白，包括宿主细胞入侵、运动、中枢代谢、内毒素的生物合成、双组分调节系统和脂肪酸代谢。我们的结果是首次对任何生物体的转录后调控进行全球分析，表明翻译水平的调控是普遍存在的，并在沙门氏菌的毒力调控和环境适应中发挥重要作用。
Using sample-matched transcriptomics and proteomics measurements it is now possible to begin to understand the impact of post-transcriptional regulatory programs in Enterobacteria. In bacteria post-transcriptional regulation is mediated by relatively few identified RNA-binding protein factors including CsrA, Hfq and SmpB. A mutation in any one of these three genes, csrA, hfq, and smpB, in Salmonella is attenuated for mouse virulence and unable to survive in macrophages. CsrA has a clearly defined specificity based on binding to a specific mRNA sequence to inhibit translation. However, the proteins regulated by Hfq and SmpB are not as clearly defined. Previous work identified proteins regulated by hfq using purification of the RNA-protein complex with direct sequencing of the bound RNAs and found binding to a surprisingly large number of transcripts. In this report we have used global proteomics to directly identify proteins regulated by Hfq or SmpB by comparing protein abundance in the parent and isogenic hfq or smpB mutant. From these same samples we also prepared RNA for microarray analysis to determine if alteration of protein expression was mediated post-transcriptionally. Samples were analyzed from bacteria grown under four different conditions; two laboratory conditions and two that are thought to mimic the intracellular environment. We show that mutants of hfq and smpB directly or indirectly modulate at least 20% and 4% of all possible Salmonella proteins, respectively, with limited correlation between transcription and protein expression. These proteins represent a broad spectrum of Salmonella proteins required for many biological processes including host cell invasion, motility, central metabolism, LPS biosynthesis, two-component regulatory systems, and fatty acid metabolism. Our results represent one of the first global analyses of post-transcriptional regulons in any organism and suggest that regulation at the translational level is widespread and plays an important role in virulence regulation and environmental adaptation for Salmonella.
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影响因子：
56.9
作者：
Guo, L;Lim, KB;Miller, SI
通讯作者：
Miller, SI
影响因子：
3.1
作者：
GarciadelPortillo, F;Stein, MA;Finlay, BB
通讯作者：
Finlay, BB
影响因子：
3.2
作者：
Ibanez-Ruiz, M;Robbe-Saule, V;Norel, F
通讯作者：
Norel, F
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
3.6
作者：
Farrant, JL;Sansone, A;Kroll, JS
通讯作者：
Kroll, JS