CRISPR-dependent endogenous gene regulation is required for virulence in piscine Streptococcus agalactiae.
CRISPR-dependent endogenous gene regulation is required for virulence in piscine Streptococcus agalactiae.
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DOI：
10.1080/22221751.2021.2002127
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发表时间：
2021-12
影响因子：
13.2
通讯作者：
Liu Y
中科院分区：
文献类型：
--
作者：
Dong Y;Ma K;Cao Q;Huang H;Nie M;Liu G;Jiang M;Lu C;Liu Y
成簇规则间隔回文重复序列（CRISPR）-Cas（CRISPR相关）系统是针对入侵的移动的遗传元件（例如噬菌体或外源质粒）的原核防御。除此之外，该系统已被证明在控制某些细菌病原体的毒力方面发挥重要作用。无乳链球菌菌株GD 201008 - 001是罗非鱼败血症和脑膜炎的病原体，含有单一的II型CRISPR-Cas系统，其中Cas9作为特征蛋白。在这项研究中，我们发现CRISPR的缺失显着降低了粘附，侵袭，细胞毒性和溶血，并导致鱼S.无乳菌株RNA-Seq鉴定了236个受CRISPR调控的内源基因，其中159个基因上调，77个基因下调。在这种细菌中，CRISPR引起的基因转录变化比cas9明显得多，表明CRISPR介导的内源性基因调控主要独立于cas9。随后的研究表明，由于CRISPR缺失，CovR/S双组分系统在转录上上调，这抑制了编码溶细胞毒素的cylE基因的表达，从而降低了β-溶血素/溶细胞素的活性。然而，CovR/S的上调不是Δ CRISPR减毒表型的贡献者。此外，我们证明了CRISPR能够抑制Toll样受体2（TLR2）激活脂蛋白Sag0671的表达，从而抑制先天免疫反应。这项研究表明，S.无乳链球菌在内源性转录物的调节方面表现出非凡的潜在能力，这有助于细菌的先天免疫逃避和毒力。
The clustered regularly interspaced palindromic repeats (CRISPR)-Cas (CRISPR-associated) system is a prokaryotic defence against invading mobile genetic elements, such as bacteriophages or exogenous plasmids. Beyond this, this system has been shown to play an important role in controlling the virulence of some bacterial pathogens. Streptococcus agalactiae strain GD201008-001, a causative agent of septicemia and meningitis in tilapia, contains a single type II CRISPR-Cas system with Cas9 as a signature protein. In this study, we found that the deletion of CRISPR significantly reduced adhesion, invasion, cytotoxicity and haemolysis, and caused severely attenuated virulence in the piscine S. agalactiae strain. RNA-Seq identified 236 endogenous genes regulated by CRISPR, with 159 genes upregulated and 77 genes downregulated. The resulting change in gene transcription by CRISPR was much more pronounced than that by cas9 in this bacterium, indicating CRISPR-mediated endogenous gene regulation was mostly independently of cas9. Subsequent studies showed that CovR/S two-component system was transcriptionally upregulated due to CRISPR deletion, which repressed the expression of the cylE gene coding for a cytolytic toxin, and thus decreased the activity of β-haemolysin/cytolysin. However, upregulation of CovR/S was not the contributor to the attenuation phenotype of ΔCRISPR. Further, we demonstrated that CRISPR is capable of repressing the expression of Toll-like receptor 2 (TLR2)-activating lipoprotein Sag0671 and thus dampens the innate immune response. This study revealed that the CRISPR system of S. agalactiae exhibited extraordinary potential capability in the regulation of endogenous transcripts, which contributes to bacterial innate immune evasion and virulence.
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影响因子：
6.4
作者：
Heussler GE;Cady KC;Koeppen K;Bhuju S;Stanton BA;O'Toole GA
通讯作者：
O'Toole GA
影响因子：
3.1
作者：
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通讯作者：
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影响因子：
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作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
3.1
作者：
DALRYMPLE, SA;LUCIAN, LA;MURRAY, R
通讯作者：
MURRAY, R
影响因子：
56.9
作者：
Aliprantis, AO;Yang, RB;Zychlinsky, A
通讯作者：
Zychlinsky, A