Mobilizable plasmids drive the spread of antimicrobial resistance genes and virulence genes in Klebsiella pneumoniae.
Mobilizable plasmids drive the spread of antimicrobial resistance genes and virulence genes in Klebsiella pneumoniae.
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DOI：
10.1186/s13073-023-01260-w
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发表时间：
2023-12-01
期刊：
影响因子：
12.3
通讯作者：
中科院分区：
文献类型：
--
作者：
肺炎克雷伯菌是一种常见的临床病原菌，其质粒是耐药性和毒力基因的主要载体。相对于自身传递的接合质粒，可移动质粒由于接合元件的缺陷而受到较少的关注。然而，可移动质粒对克雷伯菌抗菌药物耐药基因和毒力基因水平转移的贡献。肺炎仍不清楚。本研究对K.通过遗传学实验和基因组分析检查肺炎链球菌。克雷伯氏菌碳青霉烯类耐药（CR）质粒pHSKP 2和多药耐药（MDR）质粒pHSKP 3。pneumoniae HS 11286、克雷伯氏菌毒力质粒pRJF 293; pneumoniae RJF 293的质粒用于缀合测定以评估可移动质粒的转移能力。构建了模拟可动员质粒和转基因质粒以证实共转移模型。通过XbaI和S1核酸酶脉冲场凝胶电泳和/或全基因组测序评价质粒形态。通过连续传代培养来分析可移动质粒在接合子中的稳定性。此外，对1194株K.进行了在肺炎链球菌中的表达以研究CRISPR-Cas系统的接合元件、货物基因和靶标的分布。对克雷伯氏菌的可移动MDR质粒和毒力质粒进行了研究。研究了携带oriT但缺乏其他接合元件的肺炎链球菌。我们的研究结果表明，携带oriT但缺乏松弛酶基因的可移动MDR和毒力质粒能够与辅助接合CR质粒跨各种克雷伯氏菌和大肠杆菌菌株共转移。受体菌株的CRISPR-Cas系统不干扰可移动质粒而不是接合质粒的转移和稳定性。计算机模拟结果表明，可移动质粒携带了20%左右的获得性耐药基因和75%以上的毒力基因。肺炎。我们的工作观察到，携带oriT但缺乏与辅助CR接合质粒和可移动质粒一起转移的松弛酶基因的可移动MDR或毒力质粒逃脱了CRISPR-Cas防御，并在受体中保持稳定。这些结果强调了可移动质粒作为重要载体在K.肺炎。在线版本包含补充材料，可通过10.1186/s13073-023-01260-w获得。
Klebsiella pneumoniae is a notorious clinical pathogen and frequently carries various plasmids, which are the main carriers of antimicrobial resistance and virulence genes. In comparison to self-transmissible conjugative plasmids, mobilizable plasmids have received much less attention due to their defects in conjugative elements. However, the contribution of mobilizable plasmids to the horizontal transfer of antimicrobial resistance genes and virulence genes of K. pneumoniae remains unclear. In this study, the transfer, stability, and cargo genes of the mobilizable plasmids of K. pneumoniae were examined via genetic experiments and genomic analysis. Carbapenem-resistant (CR) plasmid pHSKP2 and multidrug-resistant (MDR) plasmid pHSKP3 of K. pneumoniae HS11286, virulence plasmid pRJF293 of K. pneumoniae RJF293 were employed in conjugation assays to assess the transfer ability of mobilizable plasmids. Mimic mobilizable plasmids and genetically modified plasmids were constructed to confirm the cotransfer models. The plasmid morphology was evaluated through XbaI and S1 nuclease pulsed-field gel electrophoresis and/or complete genome sequencing. Mobilizable plasmid stability in transconjugants was analyzed via serial passage culture. In addition, in silico genome analysis of 3923 plasmids of 1194 completely sequenced K. pneumoniae was performed to investigate the distribution of the conjugative elements, the cargo genes, and the targets of the CRISPR-Cas system. The mobilizable MDR plasmid and virulence plasmid of K. pneumoniae were investigated, which carry oriT but lack other conjugative elements. Our results showed that mobilizable MDR and virulence plasmids carrying oriT but lacking the relaxase gene were able to cotransfer with a helper conjugative CR plasmid across various Klebsiella and Escherichia coli strains. The transfer and stability of mobilizable plasmids rather than conjugative plasmids were not interfered with by the CRISPR–Cas system of recipient strains. According to the in silico analysis, the mobilizable plasmids carry about twenty percent of acquired antimicrobial resistance genes and more than seventy-five percent of virulence genes in K. pneumoniae. Our work observed that a mobilizable MDR or virulence plasmid that carries oriT but lacks the relaxase genes transferred with the helper CR conjugative plasmid and mobilizable plasmids escaped from CRISPR–Cas defence and remained stable in recipients. These results highlight the threats of mobilizable plasmids as vital vehicles in the dissemination of antibiotic resistance and virulence genes in K. pneumoniae. The online version contains supplementary material available at 10.1186/s13073-023-01260-w.
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影响因子：
16.6
作者：
Lam MMC;Wick RR;Watts SC;Cerdeira LT;Wyres KL;Holt KE
通讯作者：
Holt KE
影响因子：
14.9
作者：
Ares-Arroyo, Manuel;Coluzzi, Charles;P.C. Rocha, Eduardo
通讯作者：
P.C. Rocha, Eduardo
影响因子：
16.8
作者：
Bottery MJ;Wood AJ;Brockhurst MA
通讯作者：
Brockhurst MA
影响因子：
14.9
作者：
O'Brien FG;Yui Eto K;Murphy RJ;Fairhurst HM;Coombs GW;Grubb WB;Ramsay JP
通讯作者：
Ramsay JP
DOI：
10.1073/pnas.2003407117
发表时间：
2020-10-06
影响因子：
11.1
作者：
David S;Cohen V;Reuter S;Sheppard AE;Giani T;Parkhill J;European Survey of Carbapenemase-Producing Enterobacteriaceae (EuSCAPE) Working Group;ESCMID Study Group for Epidemiological Markers (ESGEM);Rossolini GM;Feil EJ;Grundmann H;Aanensen DM
通讯作者：
Aanensen DM