I-E型CRISPR-Cas系统调控普拉伦沙门氏菌肠炎血清中fimZY和T3SS1基因的表达。

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology Pub Date : 2024-11-15 DOI:10.1016/j.vetmic.2024.110301

Kai Zhang , Pengyu Wang , Shanshan Li , Xiaolei Xie , Zhenyu Wang , Yang Li , Xinan Jiao , Qiuchun Li

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引用次数: 0

摘要

成簇的有规则间隔短回文重复序列和相关的 Cas 蛋白（CRISPR-Cas）为原核生物提供了对抗质粒或噬菌体入侵的适应性免疫。在沙门氏菌中，I-E型CRISPR-Cas系统通常被认为在对外来遗传因子的免疫中保持沉默。为了阐明CRISPR-Cas系统的作用，我们选择了肠炎沙门氏菌（Salmonella enterica serovar Pullorum S06004）作为模式生物，因为在以前的研究中观察到了它的四个间隔和明确的生物学特征。Western 印迹分析显示，体外培养的 S06004 中表达了 Cas3，但质粒转化试验表明，野生型（WT）和过表达 LeuO（CRISPR-Cas 的正调控因子）的 S06004 菌株对目标质粒没有免疫力。与 WT 相比，RNA-Seq 分析检测到 cas 簇突变体中编码 I 型缘毛的 fim 簇和 T3SS1 相关基因显著下调。这种下调在 CR1 突变体和单个 cas 基因的 qRT-PCR 中得到了进一步证实。因此，CR1和cas基因簇突变体对鸡肝癌细胞的侵袭能力下降。T3SS1基因在S. Pullorum、S. Enteritidis和S. Typhimurium中受到CRISPR-Cas系统的一致调控，这表明I-E型CRISPR-Cas系统在促进细菌毒力方面发挥着共同的作用。然而，这种调控的具体分子机制还需要进一步研究。

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Type I-E CRISPR-Cas system regulates fimZY and T3SS1 genes expression in Salmonella enterica serovar Pullorum

Clustered regularly interspaced short palindromic repeats and associated Cas proteins (CRISPR-Cas) provide prokaryotes with adaptive immunity against invasion by plasmids or phages. In Salmonella, the type I-E CRISPR-Cas system is typically considered silent in immunity against foreign genetic elements. To elucidate the role of the CRISPR-Cas system, we chose Salmonella enterica serovar Pullorum S06004 as a model organism due to its four spacers and well-defined biological characteristics observed in previous studies. Western blot analysis revealed expression of Cas3 in S06004 cultured in vitro, but plasmid transformation assays demonstrated that both wild-type (WT) and S06004 strains overexpressing LeuO (a positive regulator of CRISPR-Cas) showed no immunity against the target plasmid. RNA-Seq analysis detected significant downregulation of the fim cluster, encoding type I fimbriae, and T3SS1-related genes in the cas cluster mutant compared to the WT. This downregulation was further confirmed in mutants of CR1 and individual cas genes by qRT-PCR. Consequently, mutants of CR1 and cas clusters exhibited decreased invasion of chicken hepatocellular carcinoma cells. The consistent regulation of T3SS1 genes by the CRISPR-Cas system in S. Pullorum, S. Enteritidis, and S. Typhimurium indicates a common role for the type I-E CRISPR-Cas system in promoting bacterial virulence. However, the specific molecular mechanisms underlying this regulation require further investigation.

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来源期刊

Veterinary microbiology 农林科学-兽医学

CiteScore

5.90

自引率

6.10%

发文量

221

审稿时长

52 days

期刊介绍： Veterinary Microbiology is concerned with microbial (bacterial, fungal, viral) diseases of domesticated vertebrate animals (livestock, companion animals, fur-bearing animals, game, poultry, fish) that supply food, other useful products or companionship. In addition, Microbial diseases of wild animals living in captivity, or as members of the feral fauna will also be considered if the infections are of interest because of their interrelation with humans (zoonoses) and/or domestic animals. Studies of antimicrobial resistance are also included, provided that the results represent a substantial advance in knowledge. Authors are strongly encouraged to read - prior to submission - the Editorials (''Scope or cope'' and ''Scope or cope II'') published previously in the journal. The Editors reserve the right to suggest submission to another journal for those papers which they feel would be more appropriate for consideration by that journal. Original research papers of high quality and novelty on aspects of control, host response, molecular biology, pathogenesis, prevention, and treatment of microbial diseases of animals are published. Papers dealing primarily with immunology, epidemiology, molecular biology and antiviral or microbial agents will only be considered if they demonstrate a clear impact on a disease. Papers focusing solely on diagnostic techniques (such as another PCR protocol or ELISA) will not be published - focus should be on a microorganism and not on a particular technique. Papers only reporting microbial sequences, transcriptomics data, or proteomics data will not be considered unless the results represent a substantial advance in knowledge. Drug trial papers will be considered if they have general application or significance. Papers on the identification of microorganisms will also be considered, but detailed taxonomic studies do not fall within the scope of the journal. Case reports will not be published, unless they have general application or contain novel aspects. Papers of geographically limited interest, which repeat what had been established elsewhere will not be considered. The readership of the journal is global.