细菌三型分泌系统效应蛋白转运的研究进展

三型分泌系统(Type 3 secretion system,T3SS)作为存在于革兰氏阴性菌中的分泌系统之一,对革兰氏阴性菌的致病有重要作用。T3SS的致病作用体现在T3SS能直接将效应蛋白转运至宿主细胞,进而通过效应蛋白调控细胞的一系列通路,促进细菌定殖于细胞。而效应蛋白的转运受到两方面因素的调控,一方面是效应蛋白本身的信号序列,另一方面是T3SS相关蛋白的辅助。本文围绕近年来T3SS的构成、效应蛋白转运机制方面的最新进展进行概要综述。     

一个细菌的致命之旅——嗜肺军团菌分泌系统及效应蛋白的研究进展

嗜肺军团菌是引起军团菌肺炎以及庞蒂亚克热的革兰氏阴性胞内病原细菌,嗜肺军团菌侵染宿主的主要特点是可以通过其IVB型毒力分泌系统,向宿主细胞内分泌超过150种的底物效应蛋白。通过这些效应蛋白的作用,嗜肺军团菌能够调整宿主细胞的胞内运输途径,改变内外环境来伪装自己的吞噬泡,干扰宿主的细胞周期,抑制宿主细胞的凋亡,从而有效逃避宿主细胞的防御功能,创造出理想的胞内增殖环境。最后,效应蛋白还可以帮助军团菌从宿主细胞中逃逸。目前,嗜肺军团菌已经成为"病原菌-宿主相互作用"的重要研究模型,其毒力分泌系统及其底物效应蛋白的功能也成为细胞微生物学的研究热点。对嗜肺军团菌分泌系统及效应蛋白的研究不仅能够帮助阐明病原细菌的致病机理,还有助于推动对宿主免疫机制的更深层次的研究。文章主要针对嗜肺军团菌的毒力分泌系统,尤其是IVB型分泌系统的结构和功能,以及底物效应蛋白的研究进展进行了综述,向读者展示出一个小小的细菌所拥有的那令人惊叹的、如此狡猾的生存策略和它精致的杀伤武器。     

嗜肺军团菌II型分泌系统及其底物的研究进展

嗜肺军团菌(Legionella pneumophila,L. pneumophila)是研究病原菌-宿主相互作用的重要模式菌株之一,其独特的分泌系统及底物效应蛋白的结构与功能是病原微生物领域的研究热点。II型分泌系统(type II secretion system,T2SS)对促进细菌在环境和人类宿主中的生存至关重要。嗜肺军团菌Legionella secretion pathway (Lsp)系统是革兰氏阴性病原菌中一个典型的T2SS。本文综述了L. pneumophila的T2SS及其底物效应蛋白的研究进展,重点介绍其结构与功能,为深入了解革兰氏阴性病原菌的T2SS功能和作用机制提供参考。     

细菌六型分泌系统的研究进展

六型分泌系统(type Ⅵ secretion system,T6SS)作为一种广泛存在于革兰氏阴性细菌中的可收缩纳米装置,通过将有毒物质,即效应因子(effector)注射于真核或原核细胞体内,杀死真核捕食者或原核竞争对手.近年来,T6SS基因的多样性、纳米装置的组装和效应因子的致病机制等都获得了广泛关注,取得了重大...     

HopAZ1, a type III effector of Pseudomonas amygdali pv. tabaci,induces a hypersensitive response in tobacco wildfire-resistant Nicotiana tabacum ‘N509’

Pseudomonas amygdali pv. tabaci (formerly Pseudomonas syringae pv. tabaci; Pta) is a gram-negative bacterium that causes bacterial wildfire disease in Nicotiana tabacum. The pathogen establishes infections by using a type III secretion system to inject type III effector proteins (T3Es) into cells, thereby interfering with the host__s immune system. To counteract the effectors, plants have evolved disease-resistance genes and mechanisms to induce strong resistance on effector recognition. By screening a series of Pta T3E-deficient mutants, we have identified HopAZ1 as the T3E that induces disease resistance in N. tabacum ‘N509’. Inoculation with the Pta ∆hopAZ1 mutant did not induce resistance to Pta in N509. We also found that the Pta ∆hopAZ1 mutant did not induce a hypersensitive response and promoted severe disease symptoms in N509. Furthermore, a C-terminal truncated HopAZ1 abolished HopAZ1-dependent cell death in N509. These results indicate that HopAZ1 is the avirulence factor that induces resistance to Pta by N509.     

Rhizobial secreted proteins as determinants of host specificity in the rhizobium–legume symbiosis

Rhizobia are Gram-negative bacteria than can elicit the formation of specialized organs, called root nodules, on leguminous host plants. Upon infection of the nodules, they differentiate into nitrogen-fixing bacteroids. An elaborate signal exchange precedes the symbiotic interaction. In general, both rhizobia and host plants exhibit narrow specificity. Rhizobial factors contributing to this specificity include Nod factors and surface polysaccharides. It is becoming increasingly clear that protein secretion is important in determining the outcome of the interaction as well. This paper discusses our current understanding of the symbiotic role played by rhizobial secreted proteins, transported both by secretion systems that are of general use, such as the type I secretion system, and by specialized, host-targeting secretion systems, such as the type III, type IV and type VI secretion systems.     

革兰氏阴性菌Ⅵ型分泌系统及其参与金属离子转运的研究进展

细菌通过其分泌系统将特定的效应蛋白输送到外界环境或进入靶细胞中,从而在细菌和宿主、细菌和微生物群落的相互作用中占据适应性优势。Ⅵ型分泌系统（The type VI secretion system,T6SS）是革兰氏阴性菌中广泛存在的大分子分泌装置,其结构和功能类似于可收缩的噬菌体尾针样,通过细胞间直接接触将细菌各种酶或毒素效应蛋白转运到原核和真核生物中,从而介导细菌间竞争以及对宿主的致病过程。有些效应蛋白还可通过非接触依赖的方式进入胞外环境来帮助细菌获取稀缺金属离子,并且它们对应激条件下细胞内金属稳态的维持至关重要。这篇综述总结了Ⅵ型分泌系统的结构、组装及其分泌的效应蛋白,并重点阐述了Ⅵ型分泌系统在多种金属离子转运机制中作用的研究进展,有助于理解T6SS在细菌间相互作用和细菌感染过程中发挥的重要作用。     

Identification of novel Xanthomonas euvesicatoria type III effector proteins by a machine‐learning approach

The Gram‐negative bacterium Xanthomonas euvesicatoria (Xcv) is the causal agent of bacterial spot disease in pepper and tomato. Xcv pathogenicity depends on a type III secretion (T3S) system that delivers effector proteins into host cells to suppress plant immunity and promote disease. The pool of known Xcv effectors includes approximately 30 proteins, most identified in the 85‐10 strain by various experimental and computational techniques. To identify additional Xcv 85‐10 effectors, we applied a genome‐wide machine‐learning approach, in which all open reading frames (ORFs) were scored according to their propensity to encode effectors. Scoring was based on a large set of features, including genomic organization, taxonomic dispersion, hypersensitive response and pathogenicity (hrp)‐dependent expression, 5′ regulatory sequences, amino acid composition bias and GC content. Thirty‐six predicted effectors were tested for translocation into plant cells using the hypersensitive response (HR)‐inducing domain of AvrBs2 as a reporter. Seven proteins (XopAU, XopAV, XopAW, XopAP, XopAX, XopAK and XopAD) harboured a functional translocation signal and their translocation relied on the HrpF translocon, indicating that they are bona fide T3S effectors. Remarkably, four belong to novel effector families. Inactivation of the xopAP gene reduced the severity of disease symptoms in infected plants. A decrease in cell death and chlorophyll content was observed in pepper leaves inoculated with the xopAP mutant when compared with the wild‐type strain. However, populations of the xopAP mutant in infected leaves were similar in size to those of wild‐type bacteria, suggesting that the reduction in virulence was not caused by impaired bacterial growth.     

细菌VI型分泌系统效应蛋白的装配及功能的研究进展

蛋白分泌作为细胞之间传递信号的途径之一,在微生物生存竞争中也扮演着重要的角色。革兰氏阴性菌可以通过Ⅵ型分泌系统(type Ⅵ secretion system, T6SS)将效应蛋白传递至胞外或原核和真核微生物中,从而介导微生物间的竞争或宿主-细菌的相互作用,最终建立竞争优势。本文主要总结了T6SS的结构与组成,并重点对效应蛋白的装配以及其与免疫蛋白的作用机制的研究进展进行阐述,为以后靶向T6SS抗菌药物的研制提供新思路。     

植物相关细菌中Ⅵ型分泌系统的功能研究进展

型分泌系统(typeⅥsecretion system,T6SS)是一种强大的细菌分子武器,它通过将效应蛋白注入原核或真核细胞而介导细菌间竞争并影响宿主的生命活动。T6SS广泛分布于革兰氏阴性菌中,主要存在于变形菌门(Proteobacteria)。尽管T6SS的研究大多集中在动物相关细菌上,但它在植物相关细菌中的作用不能被忽视。本文对植物相关细菌的T6SS进行了较为详细的介绍,主要从T6SS的发现、T6SS在植物相关细菌间竞争中的作用、在细菌与植物互作中的作用以及在植物生物防治中的作用等4个方面综述了最新的研究成果,旨在为今后更好地研究植物相关细菌T6SS的生物学功能及其应用提供指导。     

布鲁氏菌Ⅳ型分泌系统效应因子调控宿主细胞功能研究进展

布鲁氏菌(Brucella)是一种全球广泛传播的人畜共患病原体,能够感染野生动物、家养动物及人类。为了在宿主体内建立并维持慢性感染,布鲁氏菌进化出了多种策略逃逸宿主的免疫应答机制,并且在细胞内大量繁殖。其核心机制主要依赖于Ⅳ型分泌系统(type Ⅳ secretion system,T4SS)及其分泌的效应因子。T4SS通过将效应因子直接注入宿主细胞内,调控宿主细胞的多种功能,从而帮助病原体逃避免疫监控、操纵宿主细胞内环境,并促进布鲁氏菌的生存与复制。本文综述了布鲁氏菌T4SS的结构与功能,以及T4SS效应因子在调控宿主细胞功能方面的最新研究进展,探讨了布鲁氏菌如何通过调节宿主细胞信号传导途径,实现对细胞内液泡的控制,并形成有利于细菌存活的复制生态位。这些研究进展为我们更好地理解布鲁氏菌感染的致病机制提供了新见解,并有助于开发更有效的预防和治疗策略来应对布鲁氏菌感染。     

A modular toolbox for Golden‐Gate‐based plasmid assembly streamlines the generation of Ralstonia solanacearum species complex knockout strains and multi‐cassette complementation constructs

Members of the Ralstonia solanacearum species complex (Rssc) cause bacterial wilt, a devastating plant disease that affects numerous economically important crops. Like other bacterial pests, Rssc injects a cocktail of effector proteins via the bacterial type III secretion system into host cells that collectively promote disease. Given their functional relevance in disease, the identification of Rssc effectors and the investigation of their in planta function are likely to provide clues on how to generate pest‐resistant crop plants. Accordingly, molecular analysis of effector function is a focus of Rssc research. The elucidation of effector function requires corresponding gene knockout strains or strains that express the desired effector variants. The cloning of DNA constructs that facilitate the generation of such strains has hindered the investigation of Rssc effectors. To overcome these limitations, we have designed, generated and functionally validated a toolkit consisting of DNA modules that can be assembled via Golden‐Gate (GG) cloning into either desired gene knockout constructs or multi‐cassette expression constructs. The Ralstonia‐GG‐kit is compatible with a previously established toolkit that facilitates the generation of DNA constructs for in planta expression. Accordingly, cloned modules, encoding effectors of interest, can be transferred to vectors for expression in Rssc strains and plant cells. As many effector genes have been cloned in the past as GATEWAY entry vectors, we have also established a conversion vector that allows the implementation of GATEWAY entry vectors into the Ralstonia‐GG‐kit. In summary, the Ralstonia‐GG‐kit provides a valuable tool for the genetic investigation of genes encoding effectors and other Rssc genes.