In search of Brucella abortus type IV secretion substrates: screening and identification of four proteins translocated into host cells through VirB system.
In search of Brucella abortus type IV secretion substrates: screening and identification of four proteins translocated into host cells through VirB system.
复制标题
DOI：
10.1111/j.1462-5822.2011.01618.x
复制
发表时间：
2011-08
影响因子：
3.4
通讯作者：
Comerci DJ
中科院分区：
文献类型：
--
作者：
Marchesini MI;Herrmann CK;Salcedo SP;Gorvel JP;Comerci DJ
IV型分泌系统（T4SS）是一种特殊的蛋白质复合物，被许多细菌病原体用于递送破坏不同宿主细胞过程的效应分子。布鲁氏菌属是兼性细胞内病原体，能够在哺乳动物细胞内存活和复制。布鲁氏菌T4SS （VirB）是破坏溶酶体融合和创造允许复制的细胞器所必需的。VirB的一个可能作用是转运调节宿主细胞功能的效应蛋白，以促进复制细胞器的生物发生。我们假设具有真核结构域或蛋白-蛋白相互作用结构域的蛋白将是宿主细胞功能调节的良好候选者。为了识别这些候选蛋白，我们进行了一个计算机筛选，寻找具有独特特征的蛋白质。用腺苷酸环化酶报告基因检测84个潜在底物的易位。通过这种方法，我们鉴定了6种在巨噬细胞样细胞感染后被递送到真核细胞质的蛋白质，我们可以确定其中4种由BAB1_1043、BAB1_2005、BAB1_1275和BAB2_0123基因编码的蛋白质需要功能性的T4SS来递送。我们通过免疫荧光共聚焦显微镜证实了virb介导的一种底物易位，我们发现n端25个氨基酸是其进入细胞所必需的。
Type IV secretion systems (T4SS) are specialized protein complexes used by many bacterial pathogens for the delivery of effector molecules that subvert varied host cellular processes. Brucella spp. are facultative intracellular pathogens capable of survival and replication inside mammalian cells. Brucella T4SS (VirB) is essential to subvert lysosome fusion and to create an organelle permissive for replication. One possible role for VirB is to translocate effector proteins that modulate host cellular functions for the biogenesis of the replicative organelle. We hypothesized that proteins with eukaryotic domains or protein-protein interaction domains, among others, would be good candidates for modulation of host cell functions. To identify these candidates, we performed an in silico screen looking for proteins with distinctive features. Translocation of 84 potential substrates was assayed using adenylate cyclase reporter. By this approach, we identified six proteins that are delivered to the eukaryotic cytoplasm upon infection of macrophage-like cells and we could determine that four of them, encoded by genes BAB1_1043, BAB1_2005, BAB1_1275 and BAB2_0123, require a functional T4SS for their delivery. We confirmed VirB-mediated translocation of one of the substrates by immunofluorescence confocal microscopy, and we found that the N-terminal 25 amino acids are required for its delivery into cells.
登录
查看更多内容
影响因子：
5.8
作者：
Delorenzi, M;Speed, T
通讯作者：
Speed, T
影响因子：
3.1
作者：
Chain, PSG;Comerci, DJ;Garcia, E
通讯作者：
Garcia, E
影响因子：
3.4
作者：
Comerci, DJ;Martínez-Lorenzo, MJ;Ugalde, RA
通讯作者：
Ugalde, RA
影响因子：
3.6
作者：
Bardill, JP;Miller, JL;Vogel, JP
通讯作者：
Vogel, JP
DOI：
10.1084/jem.20030088
发表时间：
2003-08-18
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Celli J;de Chastellier C;Franchini DM;Pizarro-Cerda J;Moreno E;Gorvel JP
通讯作者：
Gorvel JP