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A Novel Cellular Autoaggregative Developmentally CRP Regulated Behaviour Generates Massively Chondrule-Like Formations over Surface of Old Escherichia coli K-12 Macrocolony Biofilms

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DOI: 10.4236/abb.2014.59086    3,193 Downloads   3,633 Views   Citations


How Escherichia coli bacteria develop a particular colonial, 3-D biofilm morphological pattern is still a poorly understood process. Recently, we reported a new E. coli K-12 morphotype exhibited by old macrocolonies described as volcano-like. The formative developmental process of this morphotype has been presented as a suitable experimental model for the study of 3D patterning in macrocolony biofilms. Here, we report the optical microscopy observations and genetic analysis that have unveiled the existence of a novel autoaggregative behaviour which generates massive lumpiness over the surface of the volcano-like macrocolonies. These lumpy formations are generated by the autoaggregation and strong interaction of tightly packed bacterial cells in structures with a chondrule-like appearance which give the colony’s surface its characteristic microscopic lumpy phenotype. Furthermore, they exhibit different levels of maturation from the edge to the center of the colony. Hence, its generation appears to follow a spatiotemporal program of development during the macrocolony’s morphogenesis. Interestingly, the agar’s hardness influences the morphology exhibited by these formations, with high agar concentration (1.5%, 15 g/L) suppressing its development. This new auto-aggregative E. coli’s behaviour does not require the activity of the biofilm master regulator CsgD, the adhesiveness of flagella, pili type 1, adhesin Ag43, β-1,6-N-acetyl-D-glucosamine polymer-PGA, cellulose or colanic acid, but it is under glucose repression and the control of cAMP receptor protein (CRP). The possible physiological role of these chondrule-like formations in the adaptability of the colony to different stressful environmental conditions is discussed.

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The authors declare no conflicts of interest.

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Gómez Gómez, J. and Amils, R. (2014) A Novel Cellular Autoaggregative Developmentally CRP Regulated Behaviour Generates Massively Chondrule-Like Formations over Surface of Old Escherichia coli K-12 Macrocolony Biofilms. Advances in Bioscience and Biotechnology, 5, 727-739. doi: 10.4236/abb.2014.59086.


[1] López, D., Vlamakis, H. and Kolter, R. (2010) Biofilms. Cold Spring Harbor Perspectives in Biology, 2, a000398.
[2] Beloin, C., Roux, A. and Ghigo, J.-M. (2008) Escherichia coli Biofilms. Current Topic in Microbiology and Immunology, 322, 249-289.
[3] Flemming, H-C. and Wingender, J. (2010) The Biofilm Matrix. Nature Reviews in Microbiology, 8, 623-633.
[4] Kostakioti, M., Hadjifrangiskou, M. and Hultgren, S.J. (2013) Bacterial Biofilms: Development, Dispersal, and Therapeutic Strategies in the Dawn of the Postantibiotic Era. Cold Spring Harbor Perspectives in Medicine, 3, a010306.
[5] Ben-Jacob, E., Cohen, I. and Gutnick, D.L. (1998) Cooperative Organization of Bacterial Colonies: From Genotype to Morphotype. Annual Review Microbiology, 52, 779-806.
[6] Shapiro, J.A. (1997) Multicellularity: The Rule Not the Exception. Lessons from Escherichia coli Colonies. In: Shapiro, J.A. and Dworkin, M., Eds., Bacteria as Multicellular Organisms, Oxford University Press, New York, 14-49.
[7] Serra, D.O., Richter, A.M., Klauck, G., Mika, F. and Hengge, R. (2013) Microanatomy at Cellular Resolution and Spatial Order of Physiological Differentiation in a Bacterial Biofilm. mBio, 4, e00103-13.
[8] Serra, D.O., Richter, A.M. and Hengge, R. (2013) Cellulose as an Architectural Element in Spatially Structured Escherichia coli Biofilms. Journal of Bacteriology, 195, 5540-5554.
[9] Gómez-Gómez, J.M. and Amils, R. (2014) Crowning: A Novel Escherichia coli Colonizing Behaviour Generating a Self-Organized Corona. BMC Research Notes, 7, 108.
[10] Hung, C., Zhou, Y., Pinkner, J.S., Dodson, K.W., Crowley, J.R., Heuser, J., Chapman, M.R., Hadjifrangiskou, M., Henderson, J.P. and Hultgren, S.J. (2013) Escherichia coli Biofilms Have an Organized and Complex Extracellular Matrix Structure. mBio, 4, e00645-13.
[11] Darton, N.C. (2010) Structure and Pattern in Bacterial Colonies. In: Olafsen, J., Ed., Experimental and Computational Techniques in Soft Condensed Matter Physics, Cambrige University Press, Cambrige, 279-325.
[12] Gomez-Gomez, J.M., Baquero, F. and Blazquez, J. (1996) Cyclic AMP Receptor Protein Positively Controls gyrA Transcription and Alters DNA Topology after Nutritional Upshift in Escherichia coli. Journal of Bacteriology, 178, 3331-3314.
[13] Da Re, S. and Ghigo, J.M. (2006) A CsgD-Independent Pathway for Cellulose Production and Biofilm Formation in Escherichia coli. Journal of Bacteriology, 188, 3073-3087.
[14] Kurihara, S., Suzuki, H., Oshida, M. and Benno, Y. (2011) A Novel Putrescine Importer Required for Type 1 Pili-Driven Surface Motility Induced by Extracellular Putrescine in Escherichia coli K-12. Journal of Biological Chemistry, 286, 10185-10192.
[15] Thomasson, M.K., Fontaine, F., De Lay, N. and Storz, G. (2012) A Small RNA That Regulates Motility and Biofilm Formation in Response to Changes in Nutrient Availability in Escherichia coli. Molecular Microbiology, 84, 17-35.
[16] De Lay, N. and Gottesman, S. (2009) The Crp-Activated Small Noncoding Regulatory RNA CyaR (RyeE) Links Nutritional Status to Group Behavior. Journal of Bacteriology, 191, 461-476.
[17] Chauhan, A., Sakamoto, C., Ghigo, J.M. and Beloin, C. (2013) Did I Pick the Right Colony? Pitfalls in the Study of Regulation of the Phase Variable Antigen 43 Adhesin. PLoS ONE, 8, e73568.
[18] Høyland-Kroghsbo, N.M., Mærkedahl, R.B. and Svenningsen, S.L. (2013) A Quorum-Sensing-Induced Bacteriophage Defense Mechanism. mBio, 4, e00362-12.
[19] Brombacher, E., Baratto, A., Dorel, C. and Landini, P. (2006) Gene Expression Regulation by the Curli Activator CsgD Protein: Modulation of Cellulose Biosynthesis and Control of Negative Determinants for Microbial Adhesion. Journal of Bacteriology, 188, 2027-2037.
[20] Pesavento, C., Becker, G., Sommerfeldt, N., Possling, A., Tschowri, N., Mehlis, A. and Hengge, R. (2008) Inverse Regulatory Coordination of Motility and Curli-Mediated Adhesion in Escherichia coli. Genes & Development, 22, 2434-2446.
[21] Ogasawara, H., Yamamoto, K. and Ishihama, A. (2011) Role of the Biofilm Master Regulator CsgD in Cross-Regulation between Biofilm Formation and Flagellar Synthesis. Journal of Bacteriology, 193, 2587-2597.
[22] Wang, X., Preston III, J.F. and Romeo, T. (2004) The pgaABCD Locus of Escherichia coli Promotes the Synthesis of a Polysaccharide Adhesin Required for Biofilm Formation. Journal of Bacteriology, 186, 2724-2734.
[23] Dierksen, K.P. and Trempy, J.E. (1996) Identification of a Second RcsA Protein, a Positive Regulator of Colanic Acid Capsular Polysaccharide Genes, in Escherichia coli. Journal of Bacteriology, 178, 5053-5056.
[24] Henderson, I.R., Meehan, M. and Owen, P. (1997) Antigen 43, a Phase-Variable Bipartite Outer Membrane Protein, Determines Colony Morphology and Autoaggregation in Escherichia coli K-12. FEMS Microbiology Letters, 149, 115-120.
[25] Danese, P.L., Pratt, L.A., Dove, S.L. and Kolter, R. (2000) The Outer Membrane Protein, Antigen 43, Mediates Cell-to-Cell Interactions within Escherichia coli Biofilms. Molecular Microbiology, 37, 424-432.
[26] Hasman, H., Schembri, M.A. and Klemm, P. (2000) Antigen 43 and Type 1 Fimbriae Determine Colony Morphology of Escherichia coli K-12. Journal of Bacteriology, 182, 1089-1095.
[27] Karatan, E. and Watnick, P. (2009) Signals, Regulatory Networks, and Materials That Build and Break Bacterial Biofilms. Microbiology and Molecular Biology Reviews, 73, 310-347.
[28] Botsford, J.L and Harman, J.G. (1992) Cyclic AMP in Prokaryotes. Microbiology Reviews, 56, 100-122.
[29] Jackson, D.W., Simecka, J.W. and Romeo, T. (2002) Catabolite Repression of Escherichia coli Biofilm Formation. Journal of Bacteriology, 184, 3406-3410.
[30] Shimada, T., Fujita, N., Yamamoto, K. and Ishihama, A. (2011) Novel Roles of cAMP Receptor Protein (CRP) in Regulation of Transport and Metabolism of Carbon Sources. PLoS ONE, 6, e20081.
[31] Korea, C.G., Badouraly, R., Prevost, M.C., Ghigo, J.M. and Beloin, C. (2010) Escherichia coli K-12 Possesses Multiple Cryptic but Functional Chaperone-Usher Fimbriae with Distinct Surface Specificities. Environmental Microbiology, 12, 1957-1977.
[32] Atlung, T. and Ingmer, H. (1997) H-NS: A Modulator of Environmentally Regulated Gene Expression. Molecular Microbiology, 24, 7-17.
[33] Wurpel, D.J., Beatson, S.A., Totsika, M., Petty, N.K. and Schembri, M.A. (2013) Chaperone-Usher Fimbriae of Escherichia coli. PLoS ONE, 8, e52835.
[34] Stewart, P.S. and Franklin, M.J. (2008) Physiological Heterogeneity in Biofilms. Nature Reviews in Microbiology, 6, 199-210.
[35] Wimpenny, J., Manz, W. and Szewzyk, U. (2000) Heterogeneity in Biofilms. FEMS Microbiology Reviews, 24, 661-671.
[36] Veening, J.W., Smits, W.K. and Kuipers, O.P. (2008) Bistability, Epigenetics, and Bet-Hedging in Bacteria. Annual Reviews of Microbiology, 62, 193-210.
[37] Gómez-Gómez, J.M. (2010) Aging in Bacteria, Immortality or Not—A Critical Review. Current Aging Science, 3, 198-218.
[38] Claessen, D., Rozen, D.E., Kuipers, O.P., Søgaard-Andersen, L. and van Wezel, G.P. (2014) Bacterial Solutions to Multicellularity: A Tale of Biofilms, Filaments and Fruiting Bodies. Nature Reviews Microbiology, 12, 115-124.
[39] Gómez-Gómez, J.M. (2013) Into the Life and Death: RecA a WISE Factor Working to Integrate Survival and Evolution in Escherichia coli. Advances in Bioscience and Biotechnology, 4, 442-449.
[40] Hasman, H., Chakraborty, T. and Klemm, P. (1999) Antigen-43 Mediated Autoaggregation of Escherichia coli Is Blocked by Fimbriation. Journal of Bacteriology, 181, 4834-4841.
[41] Budrene, E.O. and Berg, H.C. (1991) Complex Patterns Formed by Motile Cells of Escherichia coli. Nature, 349, 630-633.
[42] Gabig, M., Herman-Antosiewicz, A., Kwiatkowska. M, Los, M., Thomas, M.S. and Wegrzyn, G. (2002) The Cell Surface Protein Ag43 Facilitates Phage Infection of Escherichia coli in the Presence of Bile Salts and Carbohydrates. Microbiology, 148, 1533-1542.
[43] Zhang, Q. and Yan, T. (2012) Correlation of Intracellular Trehalose Concentration with Desiccation Resistance of Soil Escherichia coli Populations. Applied and Environmental Microbiology, 78, 7407-7413.

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