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Abstract:

Pseudomonas extremaustralis is a versatile Antarctic bacterium, able to grow under microaerobic and anaerobic conditions and is related to several non-pathogenic Pseudomonads. Here we report on the role of the global anaerobic regulator Anr, in the early steps of P. extremaustralis biofilm development. We found that the anr mutant was reduced in its ability to attach, to form aggregates and to display twitching motility but presented higher swimming motility than the wild type. In addition, microscopy revealed that the wild type biofilm contained more biomass and was thicker, but were less rough than that of the anr mutant. In silico analysis of the P. extremaustralis genome for Anr-like binding sites led to the identification of two biofilm-related genes as potential targets of this regulator. When measured using Quantitative Real Time PCR, we found that the anr mutant expressed lower levels of pilG, which encodes a component of Type IV pili and has been previously implicated in cellular adhesion. Levels of morA, involved in signal transduction and flagella development, were also lower in the mutant. Our data suggest that under low oxygen conditions, such as those encountered in biofilms, Anr differentially regulates aggregation and motility thus affecting the first stages of biofilm formation. © 2013 Tribelli et al.

Registro:

Documento: Artículo
Título:The Global Anaerobic Regulator Anr, Is Involved in Cell Attachment and Aggregation Influencing the First Stages of Biofilm Development in Pseudomonas extremaustralis
Autor:Tribelli, P.M.; Hay, A.G.; López, N.I.
Filiación:IQUIBICEN-CONICET and Dpto. de Química Biológica, Universidad de Buenos Aires, Buenos Aires, Argentina
Department of Microbiology, Cornell University, Ithaca, NY, United States
Palabras clave:Anr gene; article; bacterial flagellum; bacterial gene; bacterial genome; bacterial growth; bacterial phenomena and functions; binding site; biofilm; biomass; biomicroscopy; cell adhesion; cell aggregation; controlled study; gene identification; gene mutation; gene targeting; nonhuman; Pseudomonas; Pseudomonas extremaustralis; quantitative analysis; real time polymerase chain reaction; regulator gene; signal transduction; swimming motility; twitching motility; wild type; Aerobiosis; Anaerobiosis; Bacterial Adhesion; Bacterial Proteins; Base Sequence; Biofilms; DNA, Intergenic; Gene Expression Regulation, Bacterial; Genes, Bacterial; Molecular Sequence Data; Movement; Mutation; Pseudomonas
Año:2013
Volumen:8
Número:10
DOI: http://dx.doi.org/10.1371/journal.pone.0076685
Título revista:PLoS ONE
Título revista abreviado:PLoS ONE
ISSN:19326203
CODEN:POLNC
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v8_n10_p_Tribelli

Referencias:

  • Decho, A.W., Microbial biofilms in intertidal systems: an overview (2000) Cont Shelf Res, 20, pp. 1257-1273
  • Breidenstein, E.B., de la Fuente-Nunez, C., Hancock, R.E., Pseudomonas aeruginosa: all roads lead to resistance (2011) Trends Microbiol, 19, pp. 419-426
  • Romling, U., Balsalobre, C., Biofilm infections, their resilience to therapy and innovative treatment strategies (2012) J Intern Med, 272, pp. 541-561
  • Romeo, T., When the party is over: a signal for dispersal of Pseudomonas aeruginosa biofilms (2006) J Bacteriol, 188, pp. 7325-7327
  • Tribelli, P.M., López, N.I., Poly(3-hydroxybutyrate) influences biofilm formation and motility in the Antarctic novel species Pseudomonas extremaustralis under cold conditions (2011) Extremophiles, 15, pp. 541-547
  • Worlitzsch, D., Tarran, T., Ulrich, M., Schwab, U., Cekici, A., Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients (2002) J Clin Invest, 109, pp. 317-332
  • Whiteley, M., Gita Bangera, M., Bumgarner, R., Parsek, M., Teitzel, G., Gene expression in Pseudomonas aeruginosa biofilms (2001) Nature, 413, pp. 860-864
  • Williamson, K., Richards, L., Perez-Osorio, A., Pitts, B., McInnerney, K., Heterogeneity in Pseudomonas aeruginosa biofilms includes expression of ribosome hibernation factors in the antibiotic-tolerant subpopulation and hypoxia-induced stress response in the metabolically active population (2012) J Bacteriol, 194, pp. 2062-2073
  • Wen, Z.T., Burne, R.A., Functional genomics approach to identifying genes required for biofilm development by Streptococcus mutans (2002) Appl Environ Microbiol, 68, pp. 1196-1203
  • Thompson, L.S., Webb, J.S., Rice, S.A., Kjelleberg, S., The alternative sigma factor RpoN regulates the quorum sensing gene rhlI in Pseudomonas aeruginosa (2003) FEMS Microbiol Lett, 220, pp. 187-195
  • Stanley, N.R., Britton, R.A., Grossman, A.D., Lazazzera, B.A., Identification of catabolite repression as a physiological regulator of biofilm formation by Bacillus subtilis by use of DNA microarrays (2003) J Bacteriol, 185, pp. 1951-1957
  • Hong, S.H., Wang, X., Wood, T.K., Controlling biofilm formation, prophage excision and cell death by rewiring global regulator H-NS of Escherichia coli (2010) Microb Biotechnol, 3, pp. 344-356
  • Folsom, J.P., Richards, L., Pitts, B., Roe, F., Ehrlich, G.D., Physiology of Pseudomonas aeruginosa in biofilms as revealed by transcriptome analysis (2012) BMC Microbiol, 10, p. 294
  • Waite, R.D., Paccanaro, A., Papakonstantinopoulou, A., Hurst, J., Saqi, M., Clustering of Pseudomonas aeruginosa transcriptomes from planktonic cultures, developing and mature biofilms reveals distinct expression profiles (2006) BMC Gen, 7, p. 162
  • Galimand, M., Gamper, M., Zimmermann, A., Haas, D., Positive FNR-like control of anaerobic arginine degradation and nitrate respiration in Pseudomonas aeruginosa (1991) J Bacteriol, 173, pp. 1598-1606
  • Schreiber, K., Krieger, R., Benkert, B., Eschbach, M., Arai, H., The anaerobic regulatory network required for Pseudomonas aeruginosa nitrate respiration (2007) J Bacteriol, 189, pp. 4310-4314
  • Ugidos, A., Morales, G., Rial, E., Williams, H.D., Rojo, F., The coordinate regulation of multiple terminal oxidases by the Pseudomonas putida ANR global regulator (2008) Environ Microbiol, 10, pp. 1690-1702
  • Vallet-Gely, I., Sharp, J., Simon Dove, L., Local and global regulators linking anaerobiosis to cupA fimbrial gene expression in Pseudomonas aeruginosa (2007) J Bacteriol, 189, pp. 8667-8676
  • O'Callaghan, J.F., Reen, J., Adams, C., O'Gara, F., Low oxygen induces the type III secretion system in Pseudomonas aeruginosa via modulation of the small RNAs rsmZ and rsmY (2011) Microbiology, 157, pp. 3417-3428
  • O'Callaghan, J.J., Reen, F., Adams, C., Casey, P.G., Gahan, C.G., A novel host responsive sensor mediates virulence and type III secretion during Pseudomonas aeruginosa-host cell interactions (2012) Microbiology, 158, pp. 1057-1070
  • Tribelli, P.M., Nikel, P.I., Oppezzo, O.J., Lopez, N.I., Anr, the anaerobic global regulator, modulates the redox state and oxidative stress resistance in Pseudomonas extremaustralis (2013) Microbiology (UK), 159, pp. 259-268
  • Sonnleitner, E., Gonzalez, N., Sorger-Domenigg, T., Heeb, S., Richter, A., The small RNA PhrS stimulates synthesis of the Pseudomonas aeruginosa quinolone signal (2011) Mol Microbiol, 80, pp. 868-885
  • Ayub, N.D., Pettinari, M.J., Ruiz, J., Lopez, N.I., A polyhydroxybutyrate producing Pseudomonas sp. isolated from Antarctic environments with high stress resistance (2004) Curr Microbiol, 49, pp. 170-174
  • López, N., Pettinari, M.J., Stackebrandt, E., Tribelli, P.M., Pötter, M., Pseudomonas extremaustralis sp. nov. A poly(3-hydroxybutyrate) producer isolated from an Antarctic environment (2009) Curr Microbiol, 59, pp. 514-519
  • Tribelli, P.M., Di Martino, C., López, N.I., Raiger Iustman, L.J., Biofilm lifestyle enhances diesel bioremediation in the Antarctic polyhydroxyalkanoate producer Pseudomonas extremaustralis (2012) Biodegradation, 23, pp. 645-651
  • Tribelli, P.M., Méndez, B., López, N.I., The oxygen sensitive global regulator, Anr, is involved in biosynthesis of poly(3-hydroxybutyrate) in Pseudomonas extremaustralis (2010) J Mol Microbiol Biotechnol, 19, pp. 180-188
  • Tribelli, P.M., Raiger Iustman, L.J., Catone, M.V., Di Martino, C., Revale, S., Genome sequence of the polyhydroxybutyrate producer Pseudomonas extremaustralis a highly stress resistant Antarctic bacterium (2012) J Bacteriol, 194, pp. 2381-2382
  • Huisman, G.W., Wonink, E., Meima, R., Kazemier, B., Terpstra, P., Metabolism of poly(3-hydroxyalkanoates) (PHAs) by Pseudomonas oleovorans. Identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of PHA (1992) J Biol Chem, 266, pp. 2191-2198
  • O'Toole, G., Kolter, R., Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development (1998) Mol Microbiol, 30, pp. 295-304
  • Sherlock, O., Munk Vejborg, R., Klemm, P., The TibA adhesin/invasin from enterotoxigenic Escherichia coli is self recognizing and induces bacterial aggregation and biofilm formation (2005) Infection and Immunity, 73, pp. 1954-1963
  • Tremblay, J., Déziel, E., Improving the reproducibility of Pseudomonas aeruginosa swarming motility assays (2008) J Basic Microb, 48, pp. 509-515
  • Darzins, A., The pilG gene product, required for Pseudomonas aeruginosa Pilus production and twitching motility, is homologous to the enteric, single-domain response regulator CheY (1993) J Bacteriol, 175, pp. 5934-5944
  • Larionov, A., Krause, A., Miller, W., A standard curve based method for relative real time PCR data processing (2005) BMC Bioinformatics, 6, p. 62
  • Cheng, H., Walker, G.C., Succinoglycan is required for initiation and elongation of infection threads during nodulation of alfalfa by Rhizobium meliloti (1998) J Bacteriol, 180, pp. 5183-5191
  • Heydorn, A., Nielsen, A.T., Hentzer, M., Sternberg, C., Givskov, M., Quantification of biofilm structures by the novel computer program COMSTAT (2000) Microbiology, 146, pp. 2395-2407
  • Münch, R., Hiller, K., Barg, H., Heldt, D., Linz, S., PRODORIC: prokaryotic database of gene regulation (2003) Nucleic Acids Res, 31, pp. 266-269
  • Trunk, K., Benkert, B., Quäck, N., Münch, R., Scheer, M., Anaerobic adaptation in Pseudomonas aeruginosa: definition of the Anr and Dnr regulons (2010) Environ Microbiol, 12, pp. 1719-1733
  • Bertrand, J.J., West, J.T., Engel, J.N., Genetic analysis of the regulation of type IV pilus function by the Chp chemosensory system of Pseudomonas aeruginosa (2010) J Bacteriol, 192, pp. 994-1010
  • Choy, W., Zhou, L., Kiu-Choong Syn, C., Zhang, L., Swarup, S., MorA defines a new class of regulators affecting flagellar development and biofilm formation in diverse Pseudomonas species (2004) J Bacteriol, 186, pp. 7221-7228
  • Jackson, A., Gross, M.J., Daniels, E., Hampton, T., Hammond, J., Anr, and its activation by PlcH activity, in Pseudomonas aeruginosa host colonization and virulence (2013) J Bacteriol, , doi:10.1128/JB.02169-12
  • Junker, L., Peters, J.E., Hay, A.G., Global analysis of candidate genes important for fitness in a competitive biofilm using DNA-array-based transposon mapping (2006) Microbiology, 152, pp. 2233-2245
  • Newell, P.D., Yoshioka, S., Hvorecny, K.L., Monds, R.D., O'Toole, G.A., Systematic analysis of diguanylate cyclases that promote biofilm formation by Pseudomonas fluorescens Pf01 (2011) J Bacteriol, 193, pp. 4685-4698
  • Klausen, M., Heydorn, A., Ragas, P., Lambertsen, L., Aaes-Jørgensen, A., Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants (2003) Mol Microbiol, 48, pp. 1511-1524
  • Shrout, J.D., Tolker-Nielsen, T., Givskov, M., Parsek, M.R., The contribution of cell-cell signaling and motility to bacterial biofilm formation (2011) MRS Bull, 36, pp. 367-373
  • Yoon, S.S., Hennigan, R., Hilliard, J.M., Ochsner, A., Parvatiyar, K., Pseudomonas aeruginosa anaerobic respiration in biofilms: relationships to cystic fibrosis pathogenesis (2002) Dev Cell, 3, pp. 593-603
  • O'Toole, G., Gibbs, K.A., Hager, P.W., Phibbs, P.V., Kolter, R., The global carbon metabolism regulator Crc is a component of a signal transduction pathway required for biofilm development by Pseudomonas aeruginosa (2000) J Bacteriol, 182, pp. 425-431
  • Borlee, B.R., Goldman, A.D., Murakami, K., Samudrala, R., Wozniak, D.J., Pseudomonas aeruginosa uses a cyclic-di-GMP-regulated adhesin to reinforce the biofilm extracellular matrix (2010) Mol Microbiol, 75, pp. 827-842
  • Klebensberger, J., Birkenmaier, A., Geffers, R., Kjelleberg, S., Philipp, B., SiaA and SiaD are essential for inducing autoaggregation as a specific response to detergent stress in Pseudomonas aeruginosa (2009) Environ Microbiol, 11, pp. 3073-3086
  • Guttenplan, S.B., Kearns, D.B., (2013) Regulation of flagellar motility during biofilm formation, , FEMS Microbiol Rev DOI: 10.1111/1574-6976.12018

Citas:

---------- APA ----------
Tribelli, P.M., Hay, A.G. & López, N.I. (2013) . The Global Anaerobic Regulator Anr, Is Involved in Cell Attachment and Aggregation Influencing the First Stages of Biofilm Development in Pseudomonas extremaustralis. PLoS ONE, 8(10).
http://dx.doi.org/10.1371/journal.pone.0076685
---------- CHICAGO ----------
Tribelli, P.M., Hay, A.G., López, N.I. "The Global Anaerobic Regulator Anr, Is Involved in Cell Attachment and Aggregation Influencing the First Stages of Biofilm Development in Pseudomonas extremaustralis" . PLoS ONE 8, no. 10 (2013).
http://dx.doi.org/10.1371/journal.pone.0076685
---------- MLA ----------
Tribelli, P.M., Hay, A.G., López, N.I. "The Global Anaerobic Regulator Anr, Is Involved in Cell Attachment and Aggregation Influencing the First Stages of Biofilm Development in Pseudomonas extremaustralis" . PLoS ONE, vol. 8, no. 10, 2013.
http://dx.doi.org/10.1371/journal.pone.0076685
---------- VANCOUVER ----------
Tribelli, P.M., Hay, A.G., López, N.I. The Global Anaerobic Regulator Anr, Is Involved in Cell Attachment and Aggregation Influencing the First Stages of Biofilm Development in Pseudomonas extremaustralis. PLoS ONE. 2013;8(10).
http://dx.doi.org/10.1371/journal.pone.0076685