Small Noncoding RNA CjNC110 Influences Motility, Autoagglutination, AI-2 Localization, Hydrogen Peroxide Sensitivity, and Chicken Colonization in Campylobacter jejuni

Kreuder, Amanda J.; Ruddell, Brandon; Mou, Kathy T.; Hassall, Alan; Zhang, Qijing; Plummer, Paul J.

doi:10.1128/iai.00245-20

Cited by 12 publications

(48 citation statements)

References 91 publications

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“…At least 20 CJsORFs overlap annotated genes. Two CJsORFs overlap the infection-related sRNA CJnc110 (Kreuder et al, 2020). However, as regulatory targets for CJnc110 are so far not clear, CJnc110 might be a small mRNA, rather than a dual (regulatory and coding) function sRNA.…”

Section: Discussionmentioning

confidence: 99%

Complementary Ribo-seq approaches map the translatome and provide a small protein census in the foodborne pathogenCampylobacter jejuni

Froschauer

Svensson

Gelhausen

et al. 2022

Preprint

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Background: Knowing the molecules encoded by bacterial pathogens and how their expression is regulated is essential to understand how they survive, colonize, and cause disease. RNA-seq technologies that map transcriptomes have revealed a wealth of new transcripts in bacterial pathogens. However, they do not provide direct evidence or coordinates for coding potential. In particular, they miss small proteins (less than or equal to 50-100 amino acids) translated from small open reading frames (sORFs). However, this still poorly annotated component of bacterial genomes shows emerging roles in bacterial physiology and virulence. Results: Here, we present an integrated approach based on complementary ribosome profiling (Ribo-seq) techniques to map the 'translatome' of Campylobacter jejuni, the most common cause of bacterial gastroenteritis. Besides conventional Ribo-seq, we employed translation initiation site (TIS) profiling to map start codons and reveal internal sORFs. We also developed a Ribo-seq approach for mapping of translation termination sites (TTS), which revealed stop codons not apparent from the reference genome in virulence-associated loci. Our translatome map confirms translation of leaderless ORFs and leader peptides, re-annotates start or stop codons of 35 genes, and reveals isoforms generated by internal start sites. It also adds 42 novel sORFs in diverse contexts to the C. jejuni annotation, such as within small RNAs, in 5 prime untranslated regions (UTRs), or internal/out-of-frame/antisense in larger ORFs. Using epitope tagging/western blot and mass spectrometry we validated expression of almost 60 annotated and novel sORFs, including cioY, which we show encodes a conserved, 34 amino acid component of the CioAB terminal oxidase. Conclusions: Overall, we provide a blueprint for integrating several Ribo-seq approaches to refine and enrich bacterial annotations.

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Section: Discussionmentioning

confidence: 99%

Complementary Ribo-seq approaches map the translatome and provide a small protein census in the foodborne pathogenCampylobacter jejuni

Froschauer

Svensson

Gelhausen

et al. 2022

Preprint

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“…Deletion of CjNC110 resulted in decreased levels of extracellular AI-2, while intracellular levels of AI-2 were signi cantly increased. Results suggest that CjNC110 may participate in the transport of AI-2 in C. jejuni (Kreuder et al 2020). However, further research is needed to explain the details of quorum sensing machinery in C. jejuni.…”

Section: Discussionmentioning

confidence: 93%

Mutagenic strategies against luxS gene affect the early stage of biofilm formation of Campylobacter jejuni

et al. 2021

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Currently, it is clear that the luxS gene has an impact on the process of bio lm formation in Campylobacter jejuni. However, even within the species naturally occurring strains of Campylobacter lacking the luxS gene exist, which can form bio lms. In order to better understand the genetic determinants and the role of quorum sensing through the LuxS/AI-2 pathway in bio lm formation, a set of mutant/complemented strains of C. jejuni 81-176 were prepared. Additionally, the impact of the mutagenic strategy used against the luxS gene was investigated. Bio lm formation was affected by both the presence and absence of the luxS gene, and by the mutagenic strategy used. Analysis by CLSM showed that all mutant strains formed signi cantly less bio lm mass when compared to the wild-type. Interestingly, the deletion mutant (∆luxS) showed a larger decrease in bio lm mass than the substitution (•luxS) and insertional inactivated ( luxS) mutants, even though all the mutant strains lost the ability to produce autoinducer-2 molecules. Moreover, the bio lm of the ∆luxS mutant lacked the characteristic microcolonies observed in all other strains. The complementation of all mutant strains resulted in restored ability to produce AI-2, to form a complex bio lm, and to develop microcolonies at the level of the wild-type.

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“…Recently, a small noncoding RNA, CjNC110, has been identified downstream of luxS in C. jejuni [ 93 ]. CjNC110-knockout mutants showed significantly increased accumulation of intracellular AI-2 compared to WT, while extracellular AI-2 levels were reduced.…”

Section: Introductionmentioning

confidence: 99%

“…CjNC110-knockout mutants showed significantly increased accumulation of intracellular AI-2 compared to WT, while extracellular AI-2 levels were reduced. This suggested that the CjNC110 RNA modulates AI-2 detection and export [ 93 ]. However, the role of this regulatory RNA in QS regulation and AI-2 export requires further investigation.…”

Section: Introductionmentioning

confidence: 99%

Campylobacter jejuni virulence factors: update on emerging issues and trends

Tikhomirova,

McNabb,

Petterlin

et al. 2024

J Biomed Sci

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Campylobacter jejuni is a very common cause of gastroenteritis, and is frequently transmitted to humans through contaminated food products or water. Importantly, C. jejuni infections have a range of short- and long-term sequelae such as irritable bowel syndrome and Guillain Barre syndrome. C. jejuni triggers disease by employing a range of molecular strategies which enable it to colonise the gut, invade the epithelium, persist intracellularly and avoid detection by the host immune response. The objective of this review is to explore and summarise recent advances in the understanding of the C. jejuni molecular factors involved in colonisation, invasion of cells, collective quorum sensing-mediated behaviours and persistence. Understanding the mechanisms that underpin the pathogenicity of C. jejuni will enable future development of effective preventative approaches and vaccines against this pathogen.

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Small Noncoding RNA CjNC110 Influences Motility, Autoagglutination, AI-2 Localization, Hydrogen Peroxide Sensitivity, and Chicken Colonization in Campylobacter jejuni

Cited by 12 publications

References 91 publications

Complementary Ribo-seq approaches map the translatome and provide a small protein census in the foodborne pathogenCampylobacter jejuni

Complementary Ribo-seq approaches map the translatome and provide a small protein census in the foodborne pathogenCampylobacter jejuni

Mutagenic strategies against luxS gene affect the early stage of biofilm formation of Campylobacter jejuni

Campylobacter jejuni virulence factors: update on emerging issues and trends

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