The Brucella suis Type IV Secretion System Assembles in the Cell Envelope of the Heterologous Host Agrobacterium tumefaciens and Increases IncQ Plasmid pLS1 Recipient Competence

Anna Carle; Christoph H?ppner; Khaled Ahmed Aly; Qing Yuan; Amke den Dulk-Ras; Annette Vergunst; David OCallaghan; Christian Baron

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The Brucella suis Type IV Secretion System Assembles in the Cell Envelope of the Heterologous Host Agrobacterium tumefaciens and Increases IncQ Plasmid pLS1 Recipient Competence

机译：猪布鲁氏菌IV型分泌系统在异源宿主农杆菌的细胞膜中组装并提高IncQ质粒pLS1的接收能力

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Pathogenic Brucella species replicate within mammalian cells, and their type IV secretion system is essential for intracellular survival and replication. The options for biochemical studies on the Brucella secretion system are limited due to the rigidity of the cells and biosafety concerns, which preclude large-scale cell culture and fractionation. To overcome these problems, we heterologously expressed the Brucella suis virB operon in the closely related α₂-proteobacterium Agrobacterium tumefaciens and showed that the VirB proteins assembled into a complex. Eight of the twelve VirB proteins were detected in the membranes of the heterologous host with specific antisera. Cross-linking indicated protein-protein interactions similar to those in other type IV secretion systems, and the results of immunofluorescence analysis supported the formation of VirB protein complexes in the cell envelope. Production of a subset of the B. suis VirB proteins (VirB3-VirB12) in A. tumefaciens strongly increased its ability to receive IncQ plasmid pLS1 in conjugation experiments, and production of VirB1 further enhanced the conjugation efficiency. Plasmid recipient competence correlated with periplasmic leakage and the detergent sensitivity of A. tumefaciens, suggesting a weakening of the cell envelope. Heterologous expression thus permits biochemical characterization of B. suis type IV secretion system assembly.

机译：致病性 Brucella 物种在哺乳动物细胞中复制，它们的IV型分泌系统对于细胞内存活和复制至关重要。由于细胞的刚性和对生物安全性的关注，使得 Brucella 分泌系统生化研究的选择受到限制，从而无法进行大规模的细胞培养和分级分离。为了克服这些问题，我们在密切相关的α_{2 -变形杆菌根癌农杆菌中异源表达了 SuucBrucella suis virB 操纵子，并显示了VirB蛋白组装成一个复合体。在具有特异性抗血清的异源宿主的膜中检测到十二种VirB蛋白中的八种。交联表明蛋白质与蛋白质的相互作用类似于其他IV型分泌系统中的相互作用，并且免疫荧光分析的结果支持在细胞膜中形成VirB蛋白质复合物。 B的子集的产生。 Suem A中的VirB蛋白（VirB3-VirB12）。在缀合实验中，根癌农杆菌极大地增强了其接收IncQ质粒pLS1的能力，而VirB1的产生进一步提高了缀合效率。质粒受体能力与周质泄漏和 A的去污剂敏感性有关。 tumefaciens ，提示细胞被膜减弱。因此，异源表达允许 B的生化表征。 suis IV型分泌系统组装。} 展开▼

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《Infection and immunity》 |2006年第1期|共10页

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Anna Carle; Christoph H?ppner; Khaled Ahmed Aly; Qing Yuan; Amke den Dulk-Ras; Annette Vergunst; David OCallaghan; Christian Baron;
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中图分类医学免疫学;

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1. A homologue of the Agrobacterium tumefaciens VirB and Bordetella pertussis Ptl type IV secretion systems is essential for intracellular survival of Brucella suis. [J] . OCallaghan D, Cazevieille C, Allardet Servent-A, Molecular Microbiology . 1999,第6期

机译：根癌土壤杆菌VirB和百日咳博德特氏菌Ptl IV型分泌系统的同系物对于猪布鲁氏菌的细胞内存活至关重要。

2. VirB1 Orthologs from Brucella suis and pKM101 Complement Defects of the Lytic Transglycosylase Required for Efficient Type IV Secretion from Agrobacterium tumefaciens [J] . Christoph H?ppner, Zhenying Liu, Natalie Domke, Journal of bacteriology . 2004,第5期

机译：猪布鲁氏菌的VirB1直系同源物和pKM101补充了根癌土壤杆菌有效IV型分泌所需的溶菌转糖基酶的缺陷

3. Interactions between Brucella suis VirB8 and Its Homolog TraJ from the Plasmid pSB102 Underline the Dynamic Nature of Type IV Secretion Systems [J] . Gisèle Bourg, Romain Sube, David OCallaghan, Journal of bacteriology . 2009,第9期

机译：猪布鲁氏菌VirB8与其来自质粒pSB102的同源TraJ之间的相互作用强调了IV型分泌系统的动态特性

4. Localization of the Agrobacterium tumefaciens Type IV Secretion System and its role in host cell attachment. [D] . Aguilar, Julieta. 2011

机译：根癌农杆菌IV型分泌系统的定位及其在宿主细胞附着中的作用。

5. The Brucella suis Type IV Secretion System Assembles in the Cell Envelope of the Heterologous Host Agrobacterium tumefaciens and Increases IncQ Plasmid pLS1 Recipient Competence [O] . Anna Carle, Christoph Höppner, Khaled Ahmed Aly, 2015

机译：猪布鲁氏菌IV型分泌系统在异源宿主土壤杆菌的细胞膜中组装并提高IncQ质粒pLS1的接收能力

6. The Brucella suis Type IV Secretion System Assembles in the Cell Envelope of the Heterologous Host Agrobacterium tumefaciens and Increases IncQ Plasmid pLS1 Recipient Competence [O] . Carle, Anna, Höppner, Christoph, Ahmed Aly, Khaled, 2005

机译：猪布鲁氏菌IV型分泌系统在异源宿主土壤杆菌的细胞膜中组装并提高IncQ质粒pLS1的接收能力

7. Characterization of a Plasmid-Encoded Type IV Secretion System in Campylobacter jejuni 81-176 [R] . Larsen, J. 2004

机译：空肠弯曲杆菌81-176中质粒编码的IV型分泌系统的表征

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3. 抗肿瘤环肽YM-216391在异源宿主中的生物合成 [J] . 王凌云 ,蹇晓红 ,唐功利 . 军事医学 . 2013,第007期

4. 6—氨基己酸二聚体水解酶基因在异源宿主中的表达 [J] . 周泽扬 ,下坂诚 . 西南农业大学学报 . 1996,第002期

5. 猪源大肠埃希菌中质粒介导喹诺酮耐药(PMQR)基因的多样性研究 [J] . 郭晶晶 ,侯思梦 ,刘连杰 . 西北农业学报 . 2021,第011期

6. 六型分泌系统在猪源肠外致病性大肠杆菌引起宿主天然免疫过程中的作用研究 [A] . 于菲菲 . 2018

1. 抗毒素SO_1445或其编码基因在提高质粒在宿主细胞中的稳定性中的应用 [P] . 中国专利： CN106834322B . 2020.09.04

2. 抗毒素SO_1445或其编码基因在提高质粒在宿主细胞中的稳定性中的应用 [P] . 中国专利： CN106834322A . 2017-06-13

3. Increased secretion of heterologous proteins by host cells by the use of substituted promoters. [P] . 外国专利： DE3689846T2 . 1994-09-22

机译：通过使用取代的启动子，宿主细胞增加了异源蛋白的分泌。

4. Genetically engineered hepatitis c virus (hcv) nucleic acid clone, plasmid clone that houses a life-size hcv cdna, dna or rna transcribed from the full-length hcv cdna, transfected and in vitro infected host cell line with hcv, non-human animal infected with hcv, hcv viral particle, in vitro test system, hcv cell free, hcv vaccine and hcv test set and processes for identifying a cell line that is permissive and an animal that is permissive for hcv infection, to select hcv with adaptive mutations that allow higher levels of hcv replication in a permissive cell line, to infect an animal with hcv, to spread hcv in vitro, to transduce an animal susceptible to hcv infection with a heterologous gene, to produce hcv viral particles, hcv particle proteins and hcv antibodies, to separate agents capable of to modulate hcv replication, to prepare a hcv nucleic acid and to detect hcv antibodies in a biological sample from a patient. [P] . 外国专利： BR9808147A . 2000-03-28

机译：基因工程丙型肝炎病毒（hcv）核酸克隆，质粒克隆，其中包含从全长hcv cdna转录而来的真人大小的hcv cdna，dna或rna，并通过非人类动物的hcv转染和体外感染宿主细胞系被hcv感染，hcv病毒颗粒，体外测试系统，无hcv细胞，hcv疫苗和hcv测试仪的感染以及鉴定允许的细胞系和允许感染hvv的动物的过程，以选择具有以下适应性突变的hcv：允许在允许的细胞系中进行更高水平的hvv复制，以hvv感染动物，在体外传播hvv，用异源基因转导易感染hvv的动物，产生hvv病毒颗粒，hvv颗粒蛋白和hvv抗体分离能够调节hcv复制的试剂，制备hcv核酸并检测患者生物样品中的hcv抗体。

5. Polypeptides translocated into cells by the VirB/VirD4 type IV secretion system and uses thereof [P] . 外国专利： EP1454987A1 . 2004-09-08

机译：通过VirB / VirD4 IV型分泌系统转移到细胞中的多肽及其用途

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The Brucella suis Type IV Secretion System Assembles in the Cell Envelope of the Heterologous Host Agrobacterium tumefaciens and Increases IncQ Plasmid pLS1 Recipient Competence

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