Increased plasmid copy number is essential for
            <i>Yersinia</i>
            T3SS function and virulence

Wang, He; Avican, Kemal; Fahlgren, Anna; Erttmann, Saskia F.; Nuss, Aaron M.; Dersch, Petra; Fällman, Maria; Edgren, Tomas; Wolf‐Watz, Hans

doi:10.1126/science.aaf7501

Cited by 69 publications

(78 citation statements)

References 35 publications

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“…The ratio of repA transcripts to its antisense RNA copA was strongly increased compared with bacteria grown at 37°C. This transcriptional profile indicated that the replication of pYV is increased during Peyer's patch colonization and supported a recent observation showing that Yersinia up-regulates the pYV copy number during infection (69).…”

Section: Resultssupporting

confidence: 89%

“…Consequently, the most important defense mechanism of the bacteria is to adjust to the stressful conditions and counteract neutrophil attack within fibrin clots. Accordingly, the pathogen's transcriptional response is characterized by a marked up-regulation of the antiphagocytic T3SS/Yop machinery in response to temperature and an increase of the virulence plasmid copy number, which is likely triggered by host cell contact (69,96). This defense strategy is supported by the induction of radical detoxifying genes, such as hmp (Fig.…”

Section: Discussionmentioning

confidence: 99%

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Tissue dual RNA-seq allows fast discovery of infection-specific functions and riboregulators shaping host–pathogen transcriptomes

Nuss

Beckstette

Пименова

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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137

117

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Pathogenic bacteria need to rapidly adjust their virulence and fitness program to prevent eradication by the host. So far, underlying adaptation processes that drive pathogenesis have mostly been studied in vitro, neglecting the true complexity of host-induced stimuli acting on the invading pathogen. In this study, we developed an unbiased experimental approach that allows simultaneous monitoring of genome-wide infection-linked transcriptional alterations of the host and colonizing extracellular pathogens. Using this tool for Yersinia pseudotuberculosis-infected lymphatic tissues, we revealed numerous alterations of host transcripts associated with inflammatory and acute-phase responses, coagulative activities, and transition metal ion sequestration, highlighting that the immune response is dominated by infiltrating neutrophils and elicits a mixed T H 17/T H 1 response. In consequence, the pathogen's response is mainly directed to prevent phagocytic attacks. Yersinia up-regulates the gene and expression dose of the antiphagocytic type III secretion system (T3SS) and induces functions counteracting neutrophil-induced ion deprivation, radical stress, and nutritional restraints. Several conserved bacterial riboregulators were identified that impacted this response. The strongest influence on virulence was found for the loss of the carbon storage regulator (Csr) system, which is shown to be essential for the up-regulation of the T3SS on host cell contact. In summary, our established approach provides a powerful tool for the discovery of infection-specific stimuli, induced host and pathogen responses, and underlying regulatory processes.tissue dual RNA-seq | host-pathogen interaction | host-adapted metabolism | noncoding RNAs | Yersinia

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Section: Resultssupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Tissue dual RNA-seq allows fast discovery of infection-specific functions and riboregulators shaping host–pathogen transcriptomes

Nuss

Beckstette

Пименова

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

137

117

View full text Add to dashboard Cite

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“…This variable sequencing depth challenges traditional k-mer filtering strategies based on a fixed, all-ornothing threshold. Additionally, it is critically important to recover such plasmids during sequencing, because increased copy number has been previously associated with virulence in other species like Yersinia pestis (Wang et al 2016). As expected, MHAP overlap sensitivity for the plasmid is low (26%) when repetitive k-mers are filtered via a fixed threshold.…”

Section: Adaptive Minhash K-mer Weightingmentioning

confidence: 84%

Canu: scalable and accurate long-read assembly via adaptivek-mer weighting and repeat separation

Koren

Walenz

Berlin³

et al. 2017

Genome Res.

6,105

4,185

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Long-read single-molecule sequencing has revolutionized de novo genome assembly and enabled the automated reconstruction of reference-quality genomes. However, given the relatively high error rates of such technologies, efficient and accurate assembly of large repeats and closely related haplotypes remains challenging. We address these issues with Canu, a successor of Celera Assembler that is specifically designed for noisy single-molecule sequences. Canu introduces support for nanopore sequencing, halves depth-of-coverage requirements, and improves assembly continuity while simultaneously reducing runtime by an order of magnitude on large genomes versus Celera Assembler 8.2. These advances result from new overlapping and assembly algorithms, including an adaptive overlapping strategy based on tf-idf weighted MinHash and a sparse assembly graph construction that avoids collapsing diverged repeats and haplotypes. We demonstrate that Canu can reliably assemble complete microbial genomes and near-complete eukaryotic chromosomes using either Pacific Biosciences (PacBio) or Oxford Nanopore technologies and achieves a contig NG50 of >21 Mbp on both human and Drosophila melanogaster PacBio data sets. For assembly structures that cannot be linearly represented, Canu provides graph-based assembly outputs in graphical fragment assembly (GFA) format for analysis or integration with complementary phasing and scaffolding techniques. The combination of such highly resolved assembly graphs with long-range scaffolding information promises the complete and automated assembly of complex genomes.

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“…Bioluminescent in vivo imaging (BLI) allows the detection, visual characterization, and semiquantification of bacteria during infection of mice in real time (29). This method has widely been used to monitor Y. pseudotuberculosis YPIII infections using an in vivo imaging spectrum (IVIS) for BLI analysis (30)(31)(32)(33). To allow similar studies in strain IP32953, we constructed bioluminescent variants of the wild-type strain and the ΔtatC and ΔsufI mutant strains by integration of the complete lux operon from Photorhabdus luminescens into the 16S rRNA gene using a thermosensitive plasmid, p16Slux.…”

Section: Resultsmentioning

confidence: 99%

The Tat Substrate SufI Is Critical for the Ability of Yersinia pseudotuberculosis To Cause Systemic Infection

et al. 2017

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The twin arginine translocation (Tat) system targets folded proteins across the inner membrane and is crucial for virulence in many important humanpathogenic bacteria. Tat has been shown to be required for the virulence of Yersinia pseudotuberculosis, and we recently showed that the system is critical for different virulence-related stress responses as well as for iron uptake. In this study, we wanted to address the role of the Tat substrates in in vivo virulence. Therefore, 22 genes encoding potential Tat substrates were mutated, and each mutant was evaluated in a competitive oral infection of mice. Interestingly, a ΔsufI mutant was essentially as attenuated for virulence as the Tat-deficient strain. We also verified that SufI was Tat dependent for membrane/periplasmic localization in Y. pseudotuberculosis. In vivo bioluminescent imaging of orally infected mice revealed that both the ΔsufI and ΔtatC mutants were able to colonize the cecum and Peyer's patches (PPs) and could spread to the mesenteric lymph nodes (MLNs). Importantly, at this point, neither the ΔtatC mutant nor the ΔsufI mutant was able to spread systemically, and they were gradually cleared. Immunostaining of MLNs revealed that both the ΔtatC and ΔsufI mutants were unable to spread from the initial infection foci and appeared to be contained by neutrophils, while wild-type bacteria readily spread to establish multiple foci from day 3 postinfection. Our results show that SufI alone is required for the establishment of systemic infection and is the major cause of the attenuation of the ΔtatC mutant.KEYWORDS Yersinia pseudotuberculosis, Tat pathway, virulence, SufI, mesenteric lymph nodes, neutrophils T he genus Yersinia includes three species that are pathogenic to humans: Yersinia pestis, the causative agent of plague, and the two enteropathogens Y. enterocolitica and Y. pseudotuberculosis, which normally cause a self-limiting disease with symptoms ranging from mild diarrhea, enterocolitis, septicemia, and mesenteric lymphadenitis to reactive arthritis in humans after ingestion of contaminated food or water (1). Once it is ingested, Y. pseudotuberculosis traverses the epithelial barrier through M cells and infects the associated lymphoid tissues, such as Peyer's patches (PPs) and cecal patches, and later spreads to the mesenteric lymph nodes (MLNs). Although the infection is usually self-limited in humans, the infection caused by the two enteropathogenic Yersinia species in mice readily progresses to become systemic and disseminates to the spleen and liver (2). The main virulence arsenal of Yersinia is the type III secretion system (T3SS), which is encoded by an ϳ70-kb virulence plasmid. The T3SS enables the translocation of virulence effector proteins directly into the cytosol of the target host cell, which results in the disruption of host signaling and early immune

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Increased plasmid copy number is essential for Yersinia T3SS function and virulence

Cited by 69 publications

References 35 publications

Tissue dual RNA-seq allows fast discovery of infection-specific functions and riboregulators shaping host–pathogen transcriptomes

Tissue dual RNA-seq allows fast discovery of infection-specific functions and riboregulators shaping host–pathogen transcriptomes

Canu: scalable and accurate long-read assembly via adaptivek-mer weighting and repeat separation

The Tat Substrate SufI Is Critical for the Ability of Yersinia pseudotuberculosis To Cause Systemic Infection

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