DOCSLIB.ORG

CRISPR-Based Screening of Genomic Island Excision Events in Bacteria

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
CRISPR-Based Screening of Genomic Island Excision Events in Bacteria CRISPR-based screening of genomic island excision events in bacteria Kurt Sellea,b, Todd R. Klaenhammera,b,1, and Rodolphe Barrangoua,1 aDepartment of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695; and bFunctional Genomics Graduate Program, North Carolina State University, Raleigh, NC 27695 Contributed by Todd R. Klaenhammer, May 19, 2015 (sent for review September 22, 2014) Genomic analysis of Streptococcus thermophilus revealed that RNAs (crRNAs) that guide Cas proteins for sequence-specific mobile genetic elements (MGEs) likely contributed to gene acqui- recognition and cleavage of target DNA complementary to the sition and loss during evolutionary adaptation to milk. Clustered spacer (7). CRISPR-Cas technology has applications in strain regularly interspaced short palindromic repeats–CRISPR-associated typing and detection (8–10), exploitation of natural/engineered genes (CRISPR-Cas), the adaptive immune system in bacteria, limits immunity against mobile genetic elements (11), programmable genetic diversity by targeting MGEs including bacteriophages, genome editing in diverse backgrounds (12), transcriptional con- transposons, and plasmids. CRISPR-Cas systems are widespread trol (13, 14), and manipulation of microbial populations in defined in streptococci, suggesting that the interplay between CRISPR- consortia (15). Cas systems and MGEs is one of the driving forces governing Although sequence features corresponding to CRISPR arrays genome homeostasis in this genus. To investigate the genetic out- were described previously in multiple organisms (16, 17), Strep- in comes resulting from CRISPR-Cas targeting of integrated MGEs, tococcus thermophilus was the first microbe in which the roles silico prediction revealed four genomic islands without essential of specific cas genes and CRISPR-array components were elu- genes in lengths from 8 to 102 kbp, totaling 7% of the genome. cidated (4). S. thermophilus is a nonpathogenic, thermophilic In this study, the endogenous CRISPR3 type II system was pro- Gram-positive bacterium used as a starter culture that catabo- grammed to target the four islands independently through plas- lizes lactose to lactic acid in the syntrophic production of yogurt mid-based expression of engineered CRISPR arrays. Targeting lacZ and various cheeses (18). S. thermophilus encodes up to four within the largest 102-kbp genomic island was lethal to wild-type CRISPR-Cas systems, two of which (CRISPR1 and CRISPR3) cells and resulted in a reduction of up to 2.5-log in the surviving population. Genotyping of Lac− survivors revealed variable dele- are classified as type II-A systems that are innately active in both acquisition and interference (4, 19). Accordingly, genomic tion events between the flanking insertion-sequence elements, all S thermophilus resulting in elimination of the Lac-encoding island. Chimeric inser- analysis of . and its bacteriophages established a tion sequence footprints were observed at the deletion junctions likely mechanism for phage/DNA protection in CRISPR-Cas S thermophilus after targeting all of the four genomic islands, suggesting a com- systems. Investigation of CRISPR-Cas systems in . mon mechanism of deletion via recombination between flanking led to bioinformatic analysis of spacer origin (4, 20), discovery of insertion sequences. These results established that self-targeting the proto-spacer adjacent motif (PAM) sequences (19, 21), un- CRISPR-Cas systems may direct significant evolution of bacterial derstanding of phage–host dynamics (22, 23), demonstration genomes on a population level, influencing genome homeostasis of Cas9 endonuclease activity (7, 24, 25), and, recently, deter- and remodeling. mination of the transactivating crRNA tracrRNA structural motifs governing function and orthogonality of type II systems CRISPR | lactic acid bacteria | transposons | IS-elements | Cas9 (26). Genomic analysis of S. thermophilus revealed evolutionary obile genetic elements (MGEs) present bacteria with Significance Mcontinuous challenges to genomic stability, promoting evolution through horizontal gene transfer. The term “MGE” The development of Clustered regularly interspaced short pal- encompasses plasmids, bacteriophages, transposable elements, indromic repeats (CRISPR)–CRISPR-associated genes (CAS)–based genomic islands, and many other specialized genetic elements technology for targeted genome editing has revolutionized (1). MGEs encompass genes conferring high rates of dissemi- molecular biology approaches, but significant and outstanding nation, adaptive advantages to the host, and genomic stability, gaps exist for applications in bacteria, the native hosts of these leading to their nearly universal presence in bacterial genomes. adaptive immune systems. This study shows that CRISPR-Cas To cope with the permanent threat of predatory bacteriophages systems can be directed to target and delete genomic islands and selfish genetic elements, bacteria have evolved both innate that are flanked by insertion-sequence elements and devoid of and adaptive immune systems targeting exogenous genetic ele- essential genes. Naturally occurring minor subpopulations har- ments. Innate immunity includes cell-wall modification, restriction/ boring deletions in genomic islands were identified and readily modification systems, and abortive phage infection (2). Clustered isolated using CRISPR-Cas screening. Promising applications of regularly interspaced short palindromic repeats (CRISPR) and this approach can define minimal bacterial genomes, determine CRISPR-associated genes (Cas) are an adaptive immune system essential genes, and characterize genetically heterogeneous targeted against invasive genetic elements in bacteria (3). CRISPR- bacterial populations. Cas–mediated immunity relies on distinct molecular processes, Author contributions: K.S., T.R.K., and R.B. designed research; K.S. performed research; categorized as acquisition, expression, and interference (3). Ac- K.S. and T.R.K. contributed new reagents/analytic tools; K.S., T.R.K., and R.B. analyzed quisition occurs via molecular sampling of foreign genetic data; and K.S., T.R.K., and R.B. wrote the paper. “ ” elements, from which short sequences, termed spacers, are in- The authors declare no conflict of interest. tegrated in a polarized fashion into the CRISPR array (4). Freely available online through the PNAS open access option. Expression of CRISPR arrays is constitutive and inducible by 1To whom correspondence may be addressed. Email: [email protected] or promoter elements within the preceding leader sequence (5, 6). [email protected]. Interference results from a corresponding transcript that is This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. processed selectively at each repeat sequence, forming CRISPR 1073/pnas.1508525112/-/DCSupplemental. 8076–8081 | PNAS | June 30, 2015 | vol. 112 | no. 26 www.pnas.org/cgi/doi/10.1073/pnas.1508525112 Downloaded by guest on September 28, 2021 adaptation to milk through the loss of carbohydrate catabolism of related IS elements were mapped to the S. thermophilus genome and virulence genes found in pathogenic streptococci (18). (Fig. 1). The IS1193 and Sth6 families of IS elements appeared S. thermophilus also underwent significant acquisition of niche- most frequently in the genome and are commonly found in related genes, such as those encoding including cold-shock pro- Streptococcus pneumoniae and Streptococcus mutans (34). IS1191 teins, copper resistance proteins, proteinases, bacteriocins, and elements were not frequent but exhibited nearly perfect identity lactose catabolism proteins (18). Insertion sequences (ISs) are between the copies identified in the genome (Figs. S1A and highly prevalent in S. thermophilus genomes and contribute to S2A). Despite the prevalence of IS1193 elements, many of these genetic heterogeneity among strains by facilitating dissemination loci were shown to be small fragments that exhibited some of islands associated with dairy adaptation genes (18). The con- polymorphism and degeneracy, but several copies with a high comitant presence of MGEs and functional CRISPR-Cas systems level of sequence identity were present also (Figs. S1B and S2B). in S. thermophilus suggests that genome homeostasis is governed The Sth6 family exhibited considerable polymorphism and high at least in part by the interplay of these dynamic forces. Thus, degeneracy, with some copies harboring significant internal de- S. thermophilus constitutes an ideal host for investigating the genetic letions (Figs. S1C and S2C). IS1167 elements were well con- outcomes of CRISPR-Cas targeting of genomic islands. served (Figs. 1D and S2D). Based on the conservation of length CRISPR-Cas systems recently have been the subject of intense and sequence of the IS1167 and IS1191 elements of S. thermo- research in genome editing applications (12), but the evolutionary philus and their relative proximity to milk adaptation genes, we roles of most endogenous microbial systems remain unknown postulate that these conserved/high-fidelity transposons were (27). Even less is known concerning evolutionary outcomes of acquired in the genome recently. housing active CRISPR-Cas systems beyond the prevention of By combining the location of predicted essential ORFs and foreign DNA uptake (7), spacer acquisition events (4), and mu- highly similar IS elements, expendable islands were identified tation caused by chromosomal self-targeting
Load more

Recommended publications
  • Adaptive Introgression Between Anopheles Sibling Species Eliminates a Major Genomic Island but Not Reproductive Isolation
    ARTICLE Received 22 Feb 2014 | Accepted 28 May 2014 | Published 25 Jun 2014 DOI: 10.1038/ncomms5248 OPEN Adaptive introgression between Anopheles sibling species eliminates a major genomic island but not reproductive isolation Chris S. Clarkson1,*, David Weetman1,*, John Essandoh1,2, Alexander E. Yawson2,3, Gareth Maslen4, Magnus Manske4, Stuart G. Field5, Mark Webster6, Tiago Anta˜o1, Bronwyn MacInnis4, Dominic Kwiatkowski4,7 & Martin J. Donnelly1,4 Adaptive introgression can provide novel genetic variation to fuel rapid evolutionary responses, though it may be counterbalanced by potential for detrimental disruption of the recipient genomic background. We examine the extent and impact of recent introgression of a strongly selected insecticide-resistance mutation (Vgsc-1014F) located within one of two exceptionally large genomic islands of divergence separating the Anopheles gambiae species pair. Here we show that transfer of the Vgsc mutation results in homogenization of the entire genomic island region (B1.5% of the genome) between species. Despite this massive dis- ruption, introgression is clearly adaptive with a dramatic rise in frequency of Vgsc-1014F and no discernable impact on subsequent reproductive isolation between species. Our results show (1) how resilience of genomes to massive introgression can permit rapid adaptive response to anthropogenic selection and (2) that even extreme prominence of genomic islands of divergence can be an unreliable indicator of importance in speciation. 1 Department of Vector Biology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK. 2 Cape Coast Department of Entomology and Wildlife, School of Biological Science, University of Cape Coast, Cape Coast, Ghana. 3 Biotechnology and Nuclear Agriculture Research Institute, Ghana Atomic Energy Commission, PO Box LG 80, Legon, Accra, Ghana.
    [Show full text]
  • Divergence of a Genomic Island Leads to the Evolution of Melanization in a Halophyte Root Fungus
    The ISME Journal https://doi.org/10.1038/s41396-021-01023-8 ARTICLE Divergence of a genomic island leads to the evolution of melanization in a halophyte root fungus 1,2 3 4 5,6 7,8,9 Zhilin Yuan ● Irina S. Druzhinina ● John G. Gibbons ● Zhenhui Zhong ● Yves Van de Peer ● 10 11 2 1,2 12 13 Russell J. Rodriguez ● Zhongjian Liu ● Xinyu Wang ● Huanshen Wei ● Qi Wu ● Jieyu Wang ● 12 3 1,2 14,15 Guohui Shi ● Feng Cai ● Long Peng ● Francis M. Martin Received: 7 October 2020 / Revised: 9 May 2021 / Accepted: 24 May 2021 © The Author(s) 2021. This article is published with open access, corrected publication 2021 Abstract Understanding how organisms adapt to extreme living conditions is central to evolutionary biology. Dark septate endophytes (DSEs) constitute an important component of the root mycobiome and they are often able to alleviate host abiotic stresses. Here, we investigated the molecular mechanisms underlying the beneficial association between the DSE Laburnicola rhizohalophila and its host, the native halophyte Suaeda salsa, using population genomics. Based on genome-wide Fst (pairwise fixation index) and Vst analyses, which compared the variance in allele frequencies of single-nucleotide polymorphisms (SNPs) and copy number 1234567890();,: 1234567890();,: variants (CNVs), respectively, we found a high level of genetic differentiation between two populations. CNV patterns revealed population-specific expansions and contractions. Interestingly, we identified a ~20 kbp genomic island of high divergence with a strong sign of positive selection. This region contains a melanin-biosynthetic polyketide synthase gene cluster linked to six additional genes likely involved in biosynthesis, membrane trafficking, regulation, and localization of melanin.
    [Show full text]
  • Helicobacter Pylori Infection Causes Characteristic DNA Damage Patterns in Human Cells
    Report Helicobacter pylori Infection Causes Characteristic DNA Damage Patterns in Human Cells Graphical Abstract Authors Max Koeppel, Fernando Garcia-Alcalde, Frithjof Glowinski, Philipp Schlaermann, Thomas F. Meyer Correspondence [email protected] In Brief The gastric pathogen H. pylori can cause cancer in humans by compromising the genomic integrity of infected cells. Koeppel et al. show that infection affects the DNA damage response and reveal a DNA damage pattern reminiscent of genomic aberrations found in gastric tumors. Highlights Accession Numbers d H. pylori impairs the DNA repair response in normal human GSE55699 epithelial cells d Distinct genomic regions show increased susceptibility to H. pylori-induced damage d DNA damage accumulates in telomere-proximal, actively transcribed regions d Susceptible genomic regions overlap with gastric cancer genomic aberrations Koeppel et al., 2015, Cell Reports 11, 1703–1713 June 23, 2015 ª2015 The Authors http://dx.doi.org/10.1016/j.celrep.2015.05.030 Cell Reports Report Helicobacter pylori Infection Causes Characteristic DNA Damage Patterns in Human Cells Max Koeppel,1 Fernando Garcia-Alcalde,1 Frithjof Glowinski,1 Philipp Schlaermann,1 and Thomas F. Meyer1,* 1Department of Molecular Biology, Max Planck Institute for Infection Biology, Charite´ platz 1, 10117 Berlin, Germany *Correspondence: [email protected] http://dx.doi.org/10.1016/j.celrep.2015.05.030 This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). SUMMARY (Jenks et al., 2003; Touati et al., 2003). This process has been linked to reduced expression of mismatch repair genes (Kim Infection with the human pathogen Helicobacter et al., 2002) and increased expression of activation-induced cyti- pylori (H.
    [Show full text]
  • Estimation of Pathogenic Potential of an Environmental Pseudomonas
    www.nature.com/scientificreports OPEN Estimation of pathogenic potential of an environmental Pseudomonas aeruginosa isolate using comparative genomics Carola Berger1, Christian Rückert2, Jochen Blom3, Korneel Rabaey4, Jörn Kalinowski2 & Miriam A. Rosenbaum1,5* The isolation and sequencing of new strains of Pseudomonas aeruginosa created an extensive dataset of closed genomes. Many of the publicly available genomes are only used in their original publication while additional in silico information, based on comparison to previously published genomes, is not being explored. In this study, we defned and investigated the genome of the environmental isolate P. aeruginosa KRP1 and compared it to more than 100 publicly available closed P. aeruginosa genomes. By using diferent genomic island prediction programs, we could identify a total of 17 genomic islands and 8 genomic islets, marking the majority of the accessory genome that covers ~ 12% of the total genome. Based on intra-strain comparisons, we are able to predict the pathogenic potential of this environmental isolate. It shares a substantial amount of genomic information with the highly virulent PSE9 and LESB58 strains. For both of these, the increased virulence has been directly linked to their accessory genome before. Hence, the integrated use of previously published data can help to minimize expensive and time consuming wetlab work to determine the pathogenetic potential. Pseudomonas aeruginosa has been isolated from terrestrial and marine soil, fresh and salt water, sewage, plants, animals, and humans 1. For the latter habitats, it is known as an opportunistic pathogen, which usually spreads to already vulnerable patients, causing ~ 10% of all nosocomial infections in most European Union hospitals2.
    [Show full text]
  • Comprehensive Analysis of Mobile Genetic Elements in the Gut Microbiome Reveals Phylum-Level Niche-Adaptive Gene Pools
    bioRxiv preprint doi: https://doi.org/10.1101/214213; this version posted December 22, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Comprehensive analysis of mobile genetic elements in the gut microbiome 2 reveals phylum-level niche-adaptive gene pools 3 Xiaofang Jiang1,2,†, Andrew Brantley Hall2,3,†, Ramnik J. Xavier1,2,3,4, and Eric Alm1,2,5,* 4 1 Center for Microbiome Informatics and Therapeutics, Massachusetts Institute of Technology, 5 Cambridge, MA 02139, USA 6 2 Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA 7 3 Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard 8 Medical School, Boston, MA 02114, USA 9 4 Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General 10 Hospital and Harvard Medical School, Boston, MA 02114, USA 11 5 MIT Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 12 02142, USA 13 † Co-first Authors 14 * Corresponding Author bioRxiv preprint doi: https://doi.org/10.1101/214213; this version posted December 22, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 15 Abstract 16 Mobile genetic elements (MGEs) drive extensive horizontal transfer in the gut microbiome. This transfer 17 could benefit human health by conferring new metabolic capabilities to commensal microbes, or it could 18 threaten human health by spreading antibiotic resistance genes to pathogens. Despite their biological 19 importance and medical relevance, MGEs from the gut microbiome have not been systematically 20 characterized.
    [Show full text]
  • Analysis and Construction of Pathogenicity Island Regulatory Pathways in Salmonella Enterica Serovar Typhi
    Journal of Integrative Bioinformatics, 7(1):145, 2010 http://journal.imbio.de Analysis and construction of pathogenicity island regulatory pathways in Salmonella enterica serovar Typhi Su Yean Ong1 *, Fui Ling Ng1, Siti Suriawati Badai1, Anton Yuryev2, Maqsudul Alam1 1 Centre for Chemical Biology, Universiti Sains Malaysia, 1st Floor, Block B, No. 10, Persiaran Bukit Jambul, 11900 Bayan Lepas, Pulau Pinang, Malaysia 2 Ariadne Genomics Inc., 9430 Key West Avenue, Suite 113, Rockville, MD 20850, USA [email protected], [email protected], [email protected], [email protected], [email protected] Summary Signal transduction through protein-protein interactions and protein modifications are the main mechanisms controlling many biological processes. Here we described the (http://creativecommons.org/licenses/by-nc-nd/3.0/). implementation of MedScan information extraction technology and Pathway Studio software (Ariadne Genomics Inc.) to create a Salmonella specific molecular interaction License database. Using the database, we have constructed several signal transduction pathways in Salmonella enterica serovar Typhi which causes Typhoid Fever, a major health threat especially in developing countries. S. Typhi has several pathogenicity islands that control Unported rapid switching between different phenotypes including adhesion and colonization, 3.0 invasion, intracellular survival, proliferation, and biofilm formation in response to environmental changes. Understanding of the detailed mechanism for S. Typhi survival in host cells is necessary for development of efficient detection and treatment of this pathogen. The constructed pathways were validated using publically available gene expression microarray data for Salmonella. Bioinformatics. 1 Introduction Integrative S. Typhi is able to survive a variety of harsh conditions and defense mechanisms existing in of the human gastrointestinal tract.
    [Show full text]
  • Identification of a Pathogenicity Island, Which Contains Genes For
    Proc. Natl. Acad. Sci. USA Vol. 96, pp. 10875–10880, September 1999 Microbiology Identification of a pathogenicity island, which contains genes for virulence and avirulence, on a large native plasmid in the bean pathogen Pseudomonas syringae pathovar phaseolicola (plant disease resistance͞hypersensitive reaction͞signal transduction) ROBERT W. JACKSON*, EVANGELOS ATHANASSOPOULOS†‡,GEORGE TSIAMIS†‡,JOHN W. MANSFIELD†§,ANE SESMA¶, ʈ DAWN L. ARNOLD*, MARJORIE J. GIBBON*, JESUS MURILLO¶,JOHN D. TAYLOR , AND ALAN VIVIAN* *Department of Biological and Biomedical Sciences, University of the West of England, Coldharbor Lane, Bristol BS16 1QY, United Kingdom; †Department of Biological Sciences, Wye College, Wye, Ashford, Kent TN25 5AH, United Kingdom; ¶Escuela Te´cnica Superior de Ingenieros Agro´nomos, Universidad Pu´blica de Navarra, 31006 Pamplona, Spain; and ʈHorticulture Research International, Wellesbourne, Warwick CV35 9EF, United Kingdom Communicated by Noel T. Keen, University of California, Riverside, CA, July 7, 1999 (received for review April 2, 1999) ABSTRACT The 154-kb plasmid was cured from race 7 Certain avr genes, although recognized by their ability to strain 1449B of the phytopathogen Pseudomonas syringae pv. activate plant defenses (the HR), also may have a role in phaseolicola (Pph). Cured strains lost virulence toward bean, pathogenicity in the absence of the interacting R gene in the causing the hypersensitive reaction in previously susceptible host plant. In some cases there is a clear, qualitative effect on cultivars. Restoration of virulence was achieved by complemen- pathogenicity of mutations in avr genes, as with avrBs2 in tation with cosmid clones spanning a 30-kb region of the plasmid certain races of Xanthomonas campestris pv. vesicatoria in that contained previously identified avirulence (avr) genes avrD, pepper and avrRpm1 in P.
    [Show full text]
  • The Obscure World of Integrative and Mobilizable Elements Gérard Guédon, Virginie Libante, Charles Coluzzi, Sophie Payot-Lacroix, Nathalie Leblond-Bourget
    The obscure world of integrative and mobilizable elements Gérard Guédon, Virginie Libante, Charles Coluzzi, Sophie Payot-Lacroix, Nathalie Leblond-Bourget To cite this version: Gérard Guédon, Virginie Libante, Charles Coluzzi, Sophie Payot-Lacroix, Nathalie Leblond-Bourget. The obscure world of integrative and mobilizable elements: Highly widespread elements that pirate bacterial conjugative systems. Genes, MDPI, 2017, 8 (11), pp.337. 10.3390/genes8110337. hal- 01686871 HAL Id: hal-01686871 https://hal.archives-ouvertes.fr/hal-01686871 Submitted on 26 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License G C A T T A C G G C A T genes Review The Obscure World of Integrative and Mobilizable Elements, Highly Widespread Elements that Pirate Bacterial Conjugative Systems Gérard Guédon *, Virginie Libante, Charles Coluzzi, Sophie Payot and Nathalie Leblond-Bourget * ID DynAMic, Université de Lorraine, INRA, 54506 Vandœuvre-lès-Nancy, France; [email protected] (V.L.); [email protected] (C.C.); [email protected] (S.P.) * Correspondence: [email protected] (G.G.); [email protected] (N.L.-B.); Tel.: +33-037-274-5142 (G.G.); +33-037-274-5146 (N.L.-B.) Received: 12 October 2017; Accepted: 15 November 2017; Published: 22 November 2017 Abstract: Conjugation is a key mechanism of bacterial evolution that involves mobile genetic elements.
    [Show full text]
  • Complete Genome Sequences of Two Enterococcus Faecium Strains and Comparative Genomic Analysis
    EXPERIMENTAL AND THERAPEUTIC MEDICINE 19: 2019-2028, 2020 Complete genome sequences of two Enterococcus faecium strains and comparative genomic analysis YONG‑QI GAN, TAO ZHANG, YONG‑QIANG GAN, ZHUANG ZHAO and BIN ZHU Guangxi Institute for Food and Drug Control, Nanning, Guangxi Zhuang Autonomous Region 530021, P.R. China Received December 1, 2018; Accepted August 12, 2019 DOI: 10.3892/etm.2020.8447 Abstract. Enterococci are used for improvement of the intes- environment when treating bacterial diarrhea (3). There are tinal environment and have clinical benefits. Enterococcus two main advantages to Enterococcus that make it the most faecalis and Enterococcus faecium have similar morpholo- popular edible probiotic in animals and humans: i) In the gies, leading to confusion between the two species. In order gastrointestinal (GI) tract, Enterococcus competes with patho- to identify the National Institute for Food and Drug Control gens and thus decreases their virulence (2); ii) Enterococcus (strain 140623) and Shin Biofermin S (strain SBS-1, one of resists acid stress and cannot be digested by GI‑secreted diges- the cocci), which are widely used clinically, the present study tive juice (4). E. faecalis and E. faecium are used as digestive sequenced and analyzed these two strains. The biochemical agents for the treatment of diarrhea caused by flatulence characteristics, gas chromatography and mass spectrometry and indigestion (2). However, these two bacteria are difficult results of 140623 and SBS-1 revealed that the two strains were to distinguish morphologically. Species identification and more similar to E. faecium than E. faecalis. The genomes of genome characterization of these strains may elucidate thera- 140623 and SBS-1 contained 2,812,926 bp and 2,797,745 bp, peutic strategies for bacterial infection and probiotic treatment respectively, based on Illumina HiSeq 2000 sequencing.
    [Show full text]
  • The Role of Integrating Conjugative Elements in Helicobacter Pylori: a Review Langgeng Agung Waskito1,2, Jeng Yih-Wu3 and Yoshio Yamaoka1,4,5*
    Waskito et al. Journal of Biomedical Science (2018) 25:86 https://doi.org/10.1186/s12929-018-0489-2 REVIEW Open Access The role of integrating conjugative elements in Helicobacter pylori: a review Langgeng Agung Waskito1,2, Jeng Yih-Wu3 and Yoshio Yamaoka1,4,5* Abstract The genome of Helicobacter pylori contains many putative genes, including a genetic region known as the Integrating Conjugative Elements of H. pylori type four secretion system (ICEHptfs). This genetic regions were originally termed as “plasticity zones/regions” due to the great genetic diversity between the original two H. pylori whole genome sequences. Upon analysis of additional genome sequences, the regions were reported to be extremely common within the genome of H. pylori. Moreover, these regions were also considered conserved rather than genetically plastic and were believed to act as mobile genetic elements transferred via conjugation. Although ICEHptfs(s) are highly conserved, these regions display great allele diversity, especially on ICEHptfs4, with three different subtypes: ICEHptfs4a, 4b, and 4c. ICEHptfs were also reported to contain a novel type 4 secretion system (T4SS) with both epidemiological and in vitro infection model studies highlighting that this novel T4SS functions primarily as a virulence factor. However, there is currently no information regarding the structure, the genes responsible for forming the T4SS, and the interaction between this T4SS and other virulence genes. Unlike the cag pathogenicity island (PAI), which contains CagA, a gene found to be essential for H. pylori virulence, these novel T4SSs have not yet been reported to contain genes that contribute significant effects to the entire system.
    [Show full text]
  • Cag, a Pathogenicity Island of Helicobacter Pylori, Encodes Type I
    Proc. Natl. Acad. Sci. USA Vol. 93, pp. 14648–14653, December 1996 Genetics cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors (secretionyinsertion sequenceyinflammationyevolution) STEFANO CENSINI,CHRISTINA LANGE,ZHAOYING XIANG*, JEAN E. CRABTREE†,PAOLO GHIARA, MARK BORODOVSKY‡,RINO RAPPUOLI, AND ANTONELLO COVACCI§ Immunobiological Research Institute of Siena, Chiron Vaccines, Via Fiorentina 1, 53100 Siena, Italy Communicated by Stanley Falkow, Stanford University School of Medicine, Stanford, CA, October 9, 1996 (received for review June 14, 1996) ABSTRACT cagA, a gene that codes for an immunodom- Genetic analysis shows that the cagA gene is present only in inant antigen, is present only in Helicobacter pylori strains that type I strains, while the vacA gene is present in both types (4). are associated with severe forms of gastroduodenal disease An active toxin is produced only by type I strains; however, the (type I strains). We found that the genetic locus that contains linkage between CagA and VacA expression is not yet clear cagA (cag) is part of a 40-kb DNA insertion that likely was since the cagA and vacA genes are located more than 300 kb acquired horizontally and integrated into the chromosomal apart on the chromosome of the H. pylori NCTC 11638 (11). glutamate racemase gene. This pathogenicity island is flanked Moreover, it has been shown that inactivation of the cagA gene by direct repeats of 31 bp. In some strains, cag is split into a has no consequence on expression of VacA or on the ability to right segment (cagI) and a left segment (cagII) by a novel induce IL-8 (12–14).
    [Show full text]
  • Whole Genome Sequence-Based Prediction of Resistance Determinants in High-Level Multidrug-Resistant Campylobacter Jejuni Isolates in Lithuania
    microorganisms Communication Whole Genome Sequence-Based Prediction of Resistance Determinants in High-Level Multidrug-Resistant Campylobacter jejuni Isolates in Lithuania Jurgita Aksomaitiene 1,*, Aleksandr Novoslavskij 1, Egle Kudirkiene 2, Ausra Gabinaitiene 1 and Mindaugas Malakauskas 1 1 Department of Food Safety and Quality, Faculty of Veterinary Medicine, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes str. 18, LT 47181 Kaunas, Lithuania; [email protected] (A.N.); [email protected] (A.G.); [email protected] (M.M.) 2 Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 4, 1870 Frederiksberg C, Denmark; [email protected] * Correspondence: [email protected] Abstract: Spread of antibiotic resistance via mobile genetic elements associates with transfer of genes providing resistance against multiple antibiotics. Use of various comparative genomics analysis techniques enables to find intrinsic and acquired genes associated with phenotypic antimicrobial resistance (AMR) in Campylobacter jejuni genome sequences with exceptionally high-level multidrug resistance. In this study, we used whole genome sequences of seven C. jejuni to identify isolate-specific genomic features associated with resistance and virulence determinants and their role in multidrug resistance (MDR). All isolates were phenotypically highly resistant to tetracycline, ciprofloxacin, and ceftriaxone (MIC range from 64 to ≥256 µg/mL). Besides, two C. jejuni isolates were resistant to gentamicin, and one was resistant to erythromycin. The extensive drug-resistance profiles were Citation: Aksomaitiene, J.; confirmed for the two C. jejuni isolates assigned to ST-4447 (CC179). The most occurring genetic Novoslavskij, A.; Kudirkiene, E.; antimicrobial-resistance determinants were tetO, beta-lactamase, and multidrug efflux pumps.
    [Show full text]