Structure-guided disruption of the pseudopilus tip complex inhibits the Type II secretion in Pseudomonas aeruginosa

Zhang, Yichen; Faucher, Frédérick; Zhang, Wenwen; Wang, Shu; Neville, Nolan; Poole, Keith; Zheng, Jun; Jia, Zongchao

doi:10.1371/journal.ppat.1007343

Cited by 21 publications

(24 citation statements)

References 78 publications

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“…However, it is noted that the packing was tight, based on the side view ( Figure S1, left panel), owing to the small c side. As shown by our previously reported results [43], the long rod-like crystals were obtained via high-concentration CT digestion. The crystals belonged to a P2 1 2 1 2 1 orthorhombic lattice, with cell dimensions of a = 61.54, b = 76.76, and c = 102.86 Å ( Figure 3B, right panel).…”

Section: Different Crystal Forms Of the Xcpvwx Complex Demonstrate DIsupporting

confidence: 66%

In Situ Proteolysis Condition-Induced Crystallization of the XcpVWX Complex in Different Lattices

Zhang

Wang

Jia

2020

IJMS

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Although prevalent in the determination of protein structures; crystallography always has the bottleneck of obtaining high-quality protein crystals for characterizing a wide range of proteins; especially large protein complexes. Stable fragments or domains of proteins are more readily to crystallize; which prompts the use of in situ proteolysis to remove flexible or unstable structures for improving crystallization and crystal quality. In this work; we investigated the effects of in situ proteolysis by chymotrypsin on the crystallization of the XcpVWX complex from the Type II secretion system of Pseudomonas aeruginosa. Different proteolysis conditions were found to result in two distinct lattices in the same crystallization solution. With a shorter chymotrypsin digestion at a lower concentration; the crystals exhibited a P3 hexagonal lattice that accommodates three complex molecules in one asymmetric unit. By contrast; a longer digestion with chymotrypsin of a 10-fold higher concentration facilitated the formation of a compact P212121 orthorhombic lattice with only one complex molecule in each asymmetric unit. The molecules in the hexagonal lattice have shown high atomic displacement parameter values compared with the ones in the orthorhombic lattice. Taken together; our results clearly demonstrate that different proteolysis conditions can result in the generation of distinct lattices in the same crystallization solution; which can be exploited in order to obtain different crystal forms of a better quality

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Section: Different Crystal Forms Of the Xcpvwx Complex Demonstrate DIsupporting

confidence: 66%

In Situ Proteolysis Condition-Induced Crystallization of the XcpVWX Complex in Different Lattices

Zhang

Wang

Jia

2020

IJMS

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“…Bacterial accumulation in worm was quantified followed previous description 30 . Worms infected for duration as indicated in the text were collected and washed three times with M9 buffer containing 1mM sodium azide and 100 μg/mL of gentamicin.…”

Section: Methodsmentioning

confidence: 99%

Organism dual RNA‐seq reveals the importance of BarA/UvrY inVibrio parahaemolyticusvirulence

Zhang¹,

Xie²,

Zhang³

et al. 2020

FASEB j.

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Elucidation of host‐pathogen interaction is essential for developing effective strategies to combat bacterial infection. Dual RNA‐Seq using cultured cells or tissues/organs as the host of pathogen has emerged as a novel strategy to understand the responses concurrently from both pathogen and host at cellular level. However, bacterial infection mostly causes systematic responses from the host at organism level where the interplay is urgently to be understood but inevitably being neglected by the current practice. Here, we developed an approach that simultaneously monitor the genome‐wide infection‐linked transcriptional alterations in both pathogenic Vibrio parahaemolyticus and the infection host nematode Caenorhabditis elegans. Besides the dynamic alterations in transcriptomes of both C. elegans and V. parahaemolyticus during infection, we identify a two‐component system, BarA/UvrY, that is important for virulence in host. BarA/UvrY not only controls the virulence factors in V. parahaemolyticus including Type III and Type VI secretion systems, but also attenuates innate immune responses in C. elegans, including repression on the MAP kinase‐mediated cascades. Thus, our study exemplifies the use of dual RNA‐Seq at organism level to uncover previously unrecognized interplay between host and pathogen.

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“…Most full‐length secretin structures published to date are from the T2SS, and include those from pathogens such as V. cholerae , E. coli , Vibrio vulnificus , Aeromonas hydrophila , Klebsiella pneumoniae , and Klebsiella oxytoca . In addition, the structural characterization of the periplasmic domains of T2SS secretins as well as inner T2SS components have been instrumental in providing insight toward a mechanistic understanding of the system as well as the development of targeted inhibitors …”

Section: Assembly Of T2ss Secretinsmentioning

confidence: 99%

Bacterial secretins: Mechanisms of assembly and membrane targeting

et al. 2020

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Secretion systems are employed by bacteria to transport macromolecules across membranes without compromising their integrities. Processes including virulence, colonization, and motility are highly dependent on the secretion of effector molecules toward the immediate cellular environment, and in some cases, into the host cytoplasm. In Type II and Type III secretion systems, as well as in Type IV pili, homomultimeric complexes known as secretins form large pores in the outer bacterial membrane, and the localization and assembly of such 1 MDa molecules often relies on pilotins or accessory proteins. Significant progress has been made toward understanding details of interactions between secretins and their partner proteins using approaches ranging from bacterial genetics to cryo electron microscopy. This review provides an overview of the mode of action of pilotins and accessory proteins for T2SS, T3SS, and T4PS secretins, highlighting recent near‐atomic resolution cryo‐EM secretin complex structures and underlining the importance of these interactions for secretin functionality.

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Structure-guided disruption of the pseudopilus tip complex inhibits the Type II secretion in Pseudomonas aeruginosa

Cited by 21 publications

References 78 publications

In Situ Proteolysis Condition-Induced Crystallization of the XcpVWX Complex in Different Lattices

In Situ Proteolysis Condition-Induced Crystallization of the XcpVWX Complex in Different Lattices

Organism dual RNA‐seq reveals the importance of BarA/UvrY inVibrio parahaemolyticusvirulence

Bacterial secretins: Mechanisms of assembly and membrane targeting

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