Lrp, a global regulator, regulates the virulence of Vibrio vulnificus.
Lrp, a global regulator, regulates the virulence of Vibrio vulnificus.
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DOI：
10.1186/s12929-017-0361-9
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发表时间：
2017-08-11
影响因子：
11
通讯作者：
Hor LI
中科院分区：
文献类型：
--
作者：
Ho YC;Hung FR;Weng CH;Li WT;Chuang TH;Liu TL;Lin CY;Lo CJ;Chen CL;Chen JW;Hashimoto M;Hor LI
从临床菌株 YJ016 中偶然分离出一种创伤弧菌减毒突变体（命名为 NY303），它会导致人类严重伤口感染和败血症。该突变体在细胞毒性、在软琼脂上的迁移和对小鼠的毒力方面存在缺陷。本研究的目的是绘制该减毒突变体的突变图谱，并进一步探讨由此鉴定的基因如何参与毒力。通过二代测序确定的突变体NY303的全基因组序列与菌株YJ016的全基因组序列进行比较，以绘制突变图谱。通过分离和表征特定基因敲除突变体，鉴定了与突变体 NY303 表型相关的基因。该基因编码一个全局调节因子 Lrp。分离出 Lrp 缺陷的突变体 YH01，并进行体外、体内和离体检查，以发现受影响的毒力机制。通过比较菌株YJ016和突变体YH01的RNA-seq转录组，进一步鉴定了Lrp的靶基因。通过基因组足迹测序鉴定了Lrp结合的启动子，并通过电泳迁移率变动分析进一步检查了与毒力相关的启动子。 lrp 的突变被证明与突变体 NY303 的细胞毒性、趋化性和毒力降低有关。突变体YH01表现出与突变体NY303类似的表型，并且在小鼠体内定植和在小鼠血清中生长有缺陷，但抗吞噬能力没有缺陷。在突变体 YH01 中，分别有 596 个和 95 个基因下调和上调。许多与 MARTX 细胞毒素分泌、趋化性和铁获取相关的基因在突变体 YH01 中下调。 Lrp 基因被证明是负向自动调节的，并且 7 个下调的毒力相关基因在其启动子中被 Lrp 结合。在这些基因的启动子中鉴定出一个 14 bp 共有序列 mkCrTTkwAyTsTG，该序列可能被 Lrp 识别。 Lrp 是一种全局调节因子，参与创伤弧菌细胞毒性、趋化性和铁获取的调节。许多与这些特性相关的基因的下调可能是（至少部分）突变体 NY303 毒力丧失的原因。本文的在线版本 (doi:10.1186/s12929-017-0361-9) 包含补充材料，可供授权用户使用。
An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence. The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay. A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes. Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303. The online version of this article (doi:10.1186/s12929-017-0361-9) contains supplementary material, which is available to authorized users.
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影响因子：
3.2
作者：
Hussa, Elizabeth A.;Casanova-Torres, Angel M.;Goodrich-Blair, Heidi
通讯作者：
Goodrich-Blair, Heidi
影响因子：
6.4
作者：
Hor, LI;Chang, TT;Wang, ST
通讯作者：
Wang, ST
DOI：
10.1093/dnares/dst013
发表时间：
2013-08
期刊：
DNA research : an international journal for rapid publication of reports on genes and genomes
影响因子：
--
作者：
Chumsakul O;Nakamura K;Kurata T;Sakamoto T;Hobman JL;Ogasawara N;Oshima T;Ishikawa S
通讯作者：
Ishikawa S
影响因子：
3.2
作者：
Baek, Chang-Ho;Wang, Shifeng;Curtiss, Roy, III
通讯作者：
Curtiss, Roy, III
DOI：
10.1073/pnas.0807227105
发表时间：
2008-12-09
影响因子：
11.1
作者：
Cho, Byung-Kwan;Barrett, Christian L.;Palsson, Bernhard O.
通讯作者：
Palsson, Bernhard O.