iTRAQ-based quantitative analysis reveals the mechanism underlying the changes in physiological activity in a glutamate racemase mutant strain of Streptococcus mutans UA159.
iTRAQ-based quantitative analysis reveals the mechanism underlying the changes in physiological activity in a glutamate racemase mutant strain of Streptococcus mutans UA159.
复制标题
基于 iTRAQ 的定量分析揭示了变形链球菌 UA159 谷氨酸消旋酶突变株生理活性变化的机制。
DOI：
10.1007/s11033-020-05463-x
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发表时间：
2020
影响因子：
2.8
通讯作者：
Zhang Jian-Ying
中科院分区：
文献类型：
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作者：
Lin Jia-Cheng;Wang Xiang-Zhu;Shen Ting;Zhang Jian-Ying
变形链球菌UA 159是人类龋齿的致病菌，具有强大的致龋潜力。我们以前的研究指出，谷氨酸消旋酶（MurI）突变株（命名为S。突变体FW 1718）具有UA 159的遗传背景，表现出形态发生改变、胁迫耐受性减弱、生物膜形成能力减弱，且机制尚不清楚。在这项研究中，我们应用同量异序标签的相对和绝对定量（iTRAQ）为基础的蛋白质组学来表征的murI突变株的蛋白质组图谱，野生型菌株在化学成分确定的培养基中，阐明的机制，S。有MurI缺陷的突变体S.通过iTRAQ-偶联LC-ESI-MS/MS评估突变体FW 1718和UA 159。此外，通过Mascot、基因本体（GO）注释、蛋白质直链组簇（COG）和京都基因和基因组百科全书（KEGG）途径分析鉴定差异表达蛋白（DEP）。最后，使用检索相互作用基因/蛋白质的搜索工具（STRING）建立蛋白质-蛋白质相互作用（PPI）网络。在鉴定的1173种细菌总蛋白中，112种DEP在突变或未突变的突变株UA 159中表现出表达模式的改变。与野生型细胞相比，ΔmurI细胞中有93个蛋白的相对表达量增加（变化倍数≥ 1.2，p < 0.05），有29个蛋白的相对表达量减少（变化倍数≤ 0.833，p < 0.05）。PPI分析揭示了一个包含191条边和122个节点的DEP复杂网络。在murIknockout后显著上调的DEPs在跨越细胞壁生物合成、能量产生和DNA复制和修复的多种功能过程中发挥作用。我们在S.表明变形链球菌的细胞膜结构改变、能量代谢的重新分配和核酸机制的增强是变形链球菌致病的重要原因。mutansresponse响应to specific具体environmental环境context上下文.
Streptococcus mutansUA159 is responsible for human dental caries with robust cariogenic potential. Our previous study noted that a glutamate racemase (MurI) mutant strain (designatedS. mutansFW1718), with the hereditary background of UA159, displayed alterations of morphogenesis, attenuated stress tolerance, and weakened biofilm-forming capabilities, accompanying with unclear mechanisms. In this study, we applied isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics to characterize the proteome profiles of themurImutant strainvs.the wild-type strain in chemically defined media to elucidate the mechanisms by whichS. mutanscopes with MurI deficiency. Whole-cell proteins ofS. mutansFW1718 and UA159 were assessed by iTRAQ-coupled LC–ESI–MS/MS. Furthermore, differentially expressed proteins (DEPs) were identified by Mascot, Gene Ontology (GO) annotation, Cluster of Orthologous Groups of proteins (COG), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Finally, a protein–protein interaction (PPI) network was established using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING). Among 1173 total bacterial proteins identified, 112 DEPs exhibited altered expression patterns inS. mutansUA159 with or without themurImutation. The ΔmurI cells displayed an increase in the relative expression of 93 proteins (fold change ≥ 1.2,p< 0.05) and a decrease in 29 proteins (fold change ≤ 0.833,p< 0.05) compared with the wild-type cells. PPI analysis revealed a complex network of DEPs containing 191 edges and 122 nodes. The DEPs significantly upregulated aftermurIknockout had roles in diverse functional processes spanning cell-wall biosynthesis, energy production, and DNA replication and repair. We identified distinct variations and diverse modulators caused bymurImutation in the proteome ofS. mutans, indicating that the modification of cell membrane structure, redistribution of energy metabolism and enhanced nucleic acid machinery contributed to theS. mutansresponse to specific environmental contexts.