A structural comparison of lipopolysaccharide biosynthesis loci of Legionella pneumophila serogroup 1 strains

BMC Microbiology

Christian Lück³  Klaus Heuner²  Johan W Mouton¹  Susan Menzel³  Alexander Thürmer³  Markus Petzold¹ 

[1] Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, PO Box 9101, Nijmegen, Netherlands;Cellular Interactions of Bacterial Pathogens, Centre for Biological Security, Division 2 (ZBS2), Robert Koch-Institute, Nordufer 20, Berlin D-13353, Germany;Institute of Medical Microbiology and Hygiene, Faculty of Medicine ‘Carl Gustav Carus’, University of Technology Dresden, Fetscherstraße 74, Dresden D-01307, Germany

关键词: Monoclonal antibody typing;    Locus organization;    Lipopolysaccharide;    Legionella pneumophila;   

Others  :  1143170
DOI  :  10.1186/1471-2180-13-198

 received in 2013-04-24, accepted in 2013-08-31,  发布年份 2013

【 摘 要 】

Background

The lipopolysaccharide (LPS) is the major immuno-dominant antigen of all Legionella species including L. pneumophila. Its diversity is the basis for the classification of L. pneumophila into serogroups and monoclonal subgroups and is thought to be involved in strain specific virulence. The understanding of the genetic basis of the LPS-antigen is incomplete. Thus, we analyzed the genetic locus involved in LPS-biosynthesis of L. pneumophila serogroup 1 (Sg1) strains with the focus on strain specific gene composition.

Results

The LPS-biosynthesis loci of 14 L. pneumophila Sg1 strains comprise two distinct regions: A 15 kb region containing LPS-biosynthesis genes that can be found in all L. pneumophila strains and a Sg1-specific 18 kb region. The 15 kb region is highly conserved among Sg1 strains as reflected by high homologies of single ORFs and by a consistent ORF arrangement. In contrast, the Sg1 specific 18 kb region is variable and partially disrupted by phage related genes. We propose that the region spanning from ORF 6 to ORF 11 of the Sg1-specific region is likely involved in late LPS-modification. Due to the high variability of this small region and various combinations of single ORFs within this region a strain specific LPS-structure could be synthesized including modifications of legionaminic acid derivates.

Conclusions

Our data clearly demonstrate that the gene structure of the LPS-biosynthesis locus of L. pneumophila Sg1 strains show significant interstrain variability. These data can be used for further functional analysis of the LPS synthesis to understand pathogenesis and reactivity with monoclonal antibodies. Moreover, variable but strain specific regions can serve as basis for the development of novel genotyping assays.

【 授权许可】

   
2013 Petzold et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Pearce M, Theodoropoulos N, Mandel M, Brown E, Reed K, Cianciotto N: Legionella cardiaca sp. nov., isolated from a case of native valve endocarditis in a human heart. Int J Syst Evol Microbiol 2012, 62:2946-2954.
• [2]Rowbotham TJ: Preliminary report on the pathogenicity of Legionella pneumophila for freshwater and soil amoebae. J Clin Pathol 1980, 33:1179-1183.
• [3]Fields BS, Benson RF, Besser RE: Legionella and Legionnaires disease: 25 years of investigation. Clin Microbiol Rev 2002, 15:506-526.
• [4]Declerck P: Biofilms: the environmental playground of Legionella pneumophila. Environ Microbiol 2010, 12:557-566.
• [5]Stewart CR, Muthye V, Cianciotto NP: Legionella pneumophila persists within biofilms formed by Klebsiella pneumoniae, Flavobacterium sp., and Pseudomonas fluorescens under dynamic flow conditions. PLoS ONE 2012, 7:e50560.
• [6]Fraser DW: Legionellosis: evidence of airborne transmission. Ann NY Acad Sci 1980, 353:61-66.
• [7]Isberg RR, Tj OC, Heidtman M: The Legionella pneumophila replication vacuole: making a cosy niche inside host cells. Nat Rev Microbiol 2009, 7:13-24.
• [8]McDade JE, Shepard CC, Fraser DW, Tsai TR, Redus MA, Dowdle WR: Legionnaires’ disease: isolation of a bacterium and demonstration of its role in other respiratory disease. N Engl J Med 1977, 297:1197-1203.
• [9]Harrison TG, Afshar B, Doshi N, Fry NK, Lee JV: Distribution of Legionella pneumophila serogroups, monoclonal antibody subgroups and DNA sequence types in recent clinical and environmental isolates from England and Wales (2000–2008). Eur J Clin Microbiol Infect Dis 2009, 28:781-791.
• [10]Joseph CA, Ricketts KD, Yadav R, Patel S: Travel-associated Legionnaires’ disease in Europe in 2009. Euro Surveill 2010, 15:5-11.
• [11]Ciesielski CA, Blaser MJ, Wang WL: Serogroup specificity of Legionella pneumophila is related to lipopolysaccharide characteristics. Infect Immun 1986, 51:397-404.
• [12]Helbig JH, Jacobs E, Lück C: Legionella pneumophila urinary antigen subtyping using monoclonal antibodies as a tool for epidemiological investigations. Eur J Clin Microbiol Infect Dis 2012, 31:1673-1677.
• [13]Helbig JH, Kurtz JB, Pastoris MC, Pelaz C, Lück C: Antigenic lipopolysaccharide components of Legionella pneumophila recognized by monoclonal antibodies: possibilities and limitations for division of the species into serogroups. J Clin Microbiol 1997, 35:2841-2845.
• [14]Helbig JH, Benson RF, Pelaz C, Jacobs E, Lück C: Identification and serotyping of atypical Legionella pneumophila strains isolated from human and environmental sources. J Appl Microbiol 2007, 102:100-105.
• [15]Joly JR, Chen YY, Ramsay D: Serogrouping and subtyping of Legionella pneumophila with monoclonal antibodies. J Clin Microbiol 1983, 18:1040-1046.
• [16]Helbig JH, Bernander S, Castellani Pastoris M, Etienne J, Gaia V, Lauwers S, Lindsay D, Lück C, Marques T, Mentula S, et al.: Pan-European study on culture-proven Legionnaires’ disease: distribution of Legionella pneumophila serogroups and monoclonal subgroups. Eur J Clin Microbiol Infect Dis 2002, 21:710-716.
• [17]Knirel YA, Valvano MA: Bacterial Lipopolysaccharides: Structure, Chemical Synthesis, Biogenisis and Interaction with Host Cells. Vienna: Springer Vienna; 2011.
• [18]Knirel YA, Rietschel ET, Marre R, Zähringer U: The structure of the O-specific chain of Legionella pneumophila serogroup 1 lipopolysaccharide. Eur J Biochem 1994, 221:239-245.
• [19]Zähringer U, Knirel YA, Lindner B, Helbig JH, Sonesson A, Marre R, Rietschel ET: The lipopolysaccharide of Legionella pneumophila serogroup 1 (strain Philadelphia 1): chemical structure and biological significance. Prog Clin Biol Res 1995, 392:113-139.
• [20]Kooistra O, Herfurth L, Lüneberg E, Frosch M, Peters T, Zähringer U: Epitope mapping of the O-chain polysaccharide of Legionella pneumophila serogroup 1 lipopolysaccharide by saturation-transfer-difference NMR spectroscopy. Eur J Biochem 2002, 269:573-582.
• [21]Lüneberg E, Zetzmann N, Alber D, Knirel YA, Kooistra O, Zähringer U, Frosch M: Cloning and functional characterization of a 30 kb gene locus required for lipopolysaccharide biosynthesis in Legionella pneumophila. Int J Med Microbiol 2000, 290:37-49.
• [22]Zou CH, Knirel YA, Helbig JH, Zähringer U, Mintz CS: Molecular cloning and characterization of a locus responsible for O acetylation of the O polysaccharide of Legionella pneumophila serogroup 1 lipopolysaccharide. J Bacteriol 1999, 181:4137-4141.
• [23]Joly JR, McKinney RM, Tobin JO, Bibb WF, Watkins ID, Ramsay D: Development of a standardized subgrouping scheme for Legionella pneumophila serogroup 1 using monoclonal antibodies. J Clin Microbiol 1986, 23:768-771.
• [24]Helbig JH, Lück PC, Knirel YA, Witzleb W, Zähringer U: Molecular characterization of a virulence-associated epitope on the lipopolysaccharide of Legionella pneumophila serogroup 1. Epidemiol Infect 1995, 115:71-78.
• [25]Amemura-Maekawa J, Kikukawa K, Helbig JH, Kaneko S, Suzuki-Hashimoto A, Furuhata K, Chang B, Murai M, Ichinose M, Ohnishi M, et al.: Distribution of monoclonal antibody subgroups and sequence-based types among Legionella pneumophila serogroup 1 isolates derived from cooling tower water, bathwater, and soil in Japan. Appl Environ Microbiol 2012, 78:4263-4270.
• [26]Harrison TG, Doshi N, Fry NK, Joseph CA: Comparison of clinical and environmental isolates of Legionella pneumophila obtained in the UK over 19 years. Clin Microbiol Infect 2007, 13:78-85.
• [27]Reimer AR, Au S, Schindle S, Bernard KA: Legionella pneumophila monoclonal antibody subgroups and DNA sequence types isolated in Canada between 1981 and 2009: Laboratory Component of National Surveillance. Eur J Clin Microbiol Infect Dis 2010, 29:191-205.
• [28]D’Auria D, Jimnez-Hernndez N, Peris-Bondia F, Moya A, Latorre A: Legionella pneumophila pangenome reveals strain-specific virulence factors. BMC Genomics 2010, 11:181. BioMed Central Full Text
• [29]Glöckner G, Albert-Weissenberger C, Weinmann E, Jacobi S, Schunder E, Steinert M, Hacker J, Heuner K: Identification and characterization of a new conjugation/type IVA secretion system (trb/tra) of Legionella pneumophila Corby localized on two mobile genomic islands. Int J Med Microbiol 2008, 298:411-428.
• [30]Cazalet C, Rusniok C, Brüggemann H, Zidane N, Magnier A, Ma L, Tichit M, Jarraud S, Bouchier C, Vandenesch F, et al.: Evidence in the Legionella pneumophila genome for exploitation of host cell functions and high genome plasticity. Nat Genet 2004, 36:1165-1173.
• [31]Chien M, Morozova I, Shi S, Sheng H, Chen J, Gomez SM, Asamani G, Hill K, Nuara J, Feder M, et al.: The genomic sequence of the accidental pathogen Legionella pneumophila. Science 2004, 305:1966-1968.
• [32]Gomez-Valero L, Rusniok C, Jarraud S, Vacherie B, Rouy Z, Barbe V, Medigue C, Etienne J, Buchrieser C: Extensive recombination events and horizontal gene transfer shaped the Legionella pneumophila genomes. BMC Genomics 2011, 12:536. BioMed Central Full Text
• [33]Schroeder GN, Petty NK, Mousnier A, Harding CR, Vogrin AJ, Wee B, Fry NK, Harrison TG, Newton HJ, Thomson NR, et al.: Legionella pneumophila strain 130b possesses a unique combination of type IV secretion systems and novel Dot/Icm secretion system effector proteins. J Bacteriol 2010, 192:6001-6016.
• [34]Cazalet C, Jarraud S, Ghavi-Helm Y, Kunst F, Glaser P, Etienne J, Buchrieser C: Multigenome analysis identifies a worldwide distributed epidemic Legionella pneumophila clone that emerged within a highly diverse species. Genome Res 2008, 18:431-441.
• [35]Merault N, Rusniok C, Jarraud S, Gomez-Valero L, Cazalet C, Marin M, Brachet E, Aegerter P, Gaillard JL, Etienne J, et al.: Specific Real-Time PCR for simultaneous detection and identification of Legionella pneumophila serogroup 1 in water and clinical samples. Appl Environ Microbiol 2011, 77:1708-1717.
• [36]Glaze PA, Watson DC, Young NM, Tanner ME: Biosynthesis of CMP-N, N-diacetyllegionaminic acid from UDP-N, N -diacetylbacillosamine in Legionella pneumophila. Biochemistry 2008, 47:3272-3282.
• [37]Schoenhofen IC, McNally DJ, Vinogradov E, Whitfield D, Young NM, Dick S, Wakarchuk WW, Brisson J-R, Logan SM: Functional characterization of dehydratase/aminotransferase pairs from Helicobacter and Campylobacter: Enzymes distinguishing the pseudaminic acid and bacillosamine biosynthetic pathways. J Biol Chem 2006, 281:723-732.
• [38]Knirel YA, Shashkov AS, Tsvetkov YE, Jansson P-E, Zähringer U: 5,7-diamino-3,5,7,9-tetradeoxynon-2-ulosonic acids in bacterial glycopolymers: Chemistry and biochemistry. Adv Carbohydr Chem Biochem 2003, 58:371-417.
• [39]Lewis AL, Hensler ME, Varki A, Nizet V: The group B streptococcal sialic acid O-acetyltransferase is encoded by neuD, a conserved component of bacterial sialic acid biosynthetic gene clusters. J Biol Chem 2006, 281:11186-11192.
• [40]McNally DJ, Aubry AJ, Hui JPM, Khieu NH, Whitfield D, Ewing CP, Guerry P, Brisson J-R, Logan SM, Soo EC: Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans. J Biol Chem 2007, 282:14463-14475.
• [41]Knirel YA, Senchenkova SN, Kocharova NA, Shashkov AS, Helbig JH, Zähringer U: Identification of a homopolymer of 5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-D-glycero-D-talo-nonulosonic acid in the lipopolysaccharides of Legionella pneumophila Non-1 serogroups. Biochemistry (Mosc) 2001, 66:1035-1041.
• [42]Sahr T, Rusniok C, Dervins-Ravault D, Sismeiro O, Coppee J-Y, Buchrieser C: Deep sequencing defines the transcriptional map of L. pneumophila and identifies growth phase-dependent regulated ncRNAs implicated in virulence. RNA Biol 2012, 9:503-519.
• [43]Farhat C, Mentasti M, Jacobs E, Fry NK, Lück C: The N-Acylneuraminate cytidyltransferase gene, neuA, is heterogenous in Legionella pneumophila strains but can be used as a marker for epidemiological typing in the consensus sequence-based typing scheme. J Clin Microbiol 2011, 49:4052-4058.
• [44]Ledesma E, Camaró ML, Carbonell E, Sacristan T, Marti A, Pellicer S, Llorca J, Herrero P, Dasi MA: Subtyping of Legionella pneumophila isolates by arbitrarily primed polymerase chain reaction. Can J Microbiol 1995, 41:846-848.
• [45]Kozak NA, Benson RF, Brown E, Alexander NT, Taylor TH, Shelton BG, Fields BS: Distribution of lag-1 alleles and sequence-based types among Legionella pneumophila serogroup 1 clinical and environmental isolates in the United States. J Clin Microbiol 2009, 47:2525-2535.
• [46]Bernander S, Jacobson K, Helbig JH, Lück C, Lundholm M: A hospital-associated outbreak of Legionnaires disease caused by Legionella pneumophila serogroup 1 is characterized by stable genetic fingerprinting but variable monoclonal antibody patterns. J Clin Microbiol 2003, 41:2503-2508.
• [47]Lück C, Freier T, Steudel C, Knirel YA, Lüneberg E, Zähringer U, Helbig JH: A point mutation in the active site of Legionella pneumophila O-acetyltransferase results in modified lipopolysaccharide but does not influence virulence. Int J Med Microbiol 2001, 291:345-352.
• [48]Wagner C, Krönert C, Lück C, Jacobs E, Cianciotto NP, Helbig JH: Random mutagenesis of Legionella pneumophila reveals genes associated with lipopolysaccharide synthesis and recognition by typing monoclonal antibodies. J Appl Microbiol 2007, 103:1975-1982.
• [49]Thürmer A, Helbig JH, Jacobs E, Lück C: PCR-based ‘serotyping’ of Legionella pneumophila. J Med Microbiol 2009, 58:588-595.
• [50]Greenfield LK, Whitfield C: Synthesis of lipopolysaccharide O-antigens by ABC transporter-dependent pathways. Carbohydr Res 2012, 356:12-24.
• [51]Price MN, Huang KH, Alm EJ, Arkin AP: A novel method for accurate operon predictions in all sequenced prokaryotes. Nucleic Acids Res 2005, 33:880-892.
• [52]Kooistra O, Lüneberg E, Knirel YA, Frosch M, Zähringer U: N-Methylation in polylegionaminic acid is associated with the phase-variable epitope of Legionella pneumophila serogroup 1 lipopolysaccharide. Identification of 5-(N, N-dimethylacetimidoyl)amino and 5-acetimidoyl(N-methyl)amino-7-acetamido-3,5,7,9-tetradeoxynon-2-ulosonic acid in the O-chain polysaccharide. Eur J Biochem 2002, 269:560-572.
• [53]von Baum H, Härter G, Essig A, Lück C, Gonser T, Embacher A, Brockmann S: Preliminary report: outbreak of Legionnaires disease in the cities of Ulm and Neu-Ulm in Germany, December 2009 - January 2010. Euro Surveill 2010, 15:19472.
• [54]Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ: Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 1997, 25:3389-3402.
• [55]Lukashin A, Borodovsky M: GeneMark.hmm: new solutions for gene finding. Nucleic Acids Res 1998, 26:1107-1115.
• [56]Rutherford K, Parkhill J, Crook J, Horsnell T, Rice P, Rajandream MA, Barrell B: Artemis: sequence visualization and annotation. Bioinformatics 2000, 16:944-945.
• [57]Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schäffer AA, Yu Y-K: Protein database searches using compositionally adjusted substitution matrices. FEBS J 2005, 272:5101-5109.
• [58]Vallenet D, Engelen S, Mornico D, Cruveiller S, Fleury L, Lajus A, Rouy Z, Roche D, Salvignol G, Scarpelli C, Médigue C: MicroScope: a platform for microbial genome annotation and comparative genomics. Database 2009, 2009:Bap21.
• [59]Marchler-Bauer A, Lu S, Anderson JB, Chitsaz F, Derbyshire MK, DeWeese-Scott C, Fong JH, Geer LY, Geer RC, Gonzales NR, et al.: CDD: a Conserved Domain Database for the functional annotation of proteins. Nucleic Acids Res 2011, 39:D225-D229.
• [60]Viswanathan VK, Edelstein PH, Pope CD, Cianciotto NP: The Legionella pneumophila iraAB locus is required for iron assimilation, intracellular infection, and virulence. Infect Immun 2000, 68:1069-1079.