Flagella and pili-mediated near-surface single-cell motility mechanisms in P. aeruginosa.

Bacterial biofilms are structured multicellular communities that are responsible for a broad range of infections. Knowing how free-swimming bacteria adapt their motility mechanisms near a surface is crucial for understanding the transition from the planktonic to the biofilm phenotype. By translating microscopy movies into searchable databases of bacterial behavior and developing image-based search engines, we were able to identify fundamental appendage-specific mechanisms for the surface motility of Pseudomonas aeruginosa. Type IV pili mediate two surface motility mechanisms: horizontally oriented crawling, by which the bacterium moves lengthwise with high directional persistence, and vertically oriented walking, by which the bacterium moves with low directional persistence and high instantaneous velocity, allowing it to rapidly explore microenvironments. The flagellum mediates two additional motility mechanisms: near-surface swimming and surface-anchored spinning, which often precedes detachment from a surface. Flagella and pili interact cooperatively in a launch sequence whereby bacteria change orientation from horizontal to vertical and then detach. Vertical orientation facilitates detachment from surfaces and thereby influences biofilm morphology.

[1]  L. Burrows,et al.  Disparate Subcellular Localization Patterns of Pseudomonas aeruginosa Type IV Pilus ATPases Involved in Twitching Motility , 2005, Journal of bacteriology.

[2]  H. Swinney,et al.  Paenibacillus dendritiformis Bacterial Colony Growth Depends on Surfactant but Not on Bacterial Motion , 2009, Journal of bacteriology.

[3]  H. Swinney,et al.  Collective motion and density fluctuations in bacterial colonies , 2010, Proceedings of the National Academy of Sciences.

[4]  A. Merz,et al.  Bacterial Surface Motility: Slime Trails, Grappling Hooks and Nozzles , 2002, Current Biology.

[5]  K Bergman,et al.  Characterization of photodamage to Escherichia coli in optical traps. , 1999, Biophysical journal.

[6]  M. Sheetz,et al.  Cooperative Retraction of Bundled Type IV Pili Enables Nanonewton Force Generation , 2008, PLoS biology.

[7]  S. Kudo,et al.  Asymmetric swimming pattern of Vibrio alginolyticus cells with single polar flagella. , 2005, FEMS microbiology letters.

[8]  R. Kolter,et al.  Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development , 1998, Molecular microbiology.

[9]  J. Mattick,et al.  Quorum Sensing Is Not Required for Twitching Motility in Pseudomonas aeruginosa , 2002, Journal of bacteriology.

[10]  H. Berg,et al.  Three-dimensional tracking of motile bacteria near a solid planar surface. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Molin,et al.  Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants , 2003, Molecular microbiology.

[12]  C. Whitchurch,et al.  Roles of type IV pili, flagellum-mediated motility and extracellular DNA in the formation of mature multicellular structures in Pseudomonas aeruginosa biofilms. , 2008, Environmental microbiology.

[13]  T. Suzuki,et al.  Isolation and characterization of multiflagellate mutants of Pseudomonas aeruginosa , 1980, Journal of bacteriology.

[14]  Julien Tremblay,et al.  Self-produced extracellular stimuli modulate the Pseudomonas aeruginosa swarming motility behaviour. , 2007, Environmental microbiology.

[15]  B. Ersbøll,et al.  Experimental reproducibility in flow-chamber biofilms. , 2000, Microbiology.

[16]  J. Mattick,et al.  A re-examination of twitching motility in Pseudomonas aeruginosa. , 1999, Microbiology.

[17]  Michael P. Sheetz,et al.  Single pilus motor forces exceed 100 pN , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[18]  George M Whitesides,et al.  Swimming in circles: motion of bacteria near solid boundaries. , 2005, Biophysical journal.

[19]  B. Maier,et al.  Multiple pilus motors cooperate for persistent bacterial movement in two dimensions. , 2010, Physical review letters.

[20]  D. Grier,et al.  Methods of Digital Video Microscopy for Colloidal Studies , 1996 .

[21]  B. Maier,et al.  Bacterial motility and clustering guided by microcontact printing. , 2009, Nano letters.

[22]  X. Nassif,et al.  Type-4 pili and meningococcal adhesiveness. , 1997, Gene.

[23]  H. Berg,et al.  Torque-speed relationship of the flagellar rotary motor of Escherichia coli. , 2000, Biophysical journal.

[24]  Søren Molin,et al.  Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms , 2003, Molecular microbiology.

[25]  H. Berg Random Walks in Biology , 2018 .

[26]  D. Kaiser,et al.  Social gliding is correlated with the presence of pili in Myxococcus xanthus. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[27]  E. Greenberg,et al.  Motility of flagellated bacteria in viscous environments , 1977, Journal of bacteriology.

[28]  L. Craig,et al.  Type IV pili: paradoxes in form and function. , 2008, Current opinion in structural biology.

[29]  A. Touhami,et al.  Nanoscale Characterization and Determination of Adhesion Forces of Pseudomonas aeruginosa Pili by Using Atomic Force Microscopy , 2006, Journal of bacteriology.

[30]  D. Chopp,et al.  The impact of quorum sensing and swarming motility on Pseudomonas aeruginosa biofilm formation is nutritionally conditional , 2006, Molecular microbiology.

[31]  T. Wood,et al.  Motility influences biofilm architecture in Escherichia coli , 2006, Applied Microbiology and Biotechnology.

[32]  M. Solomon,et al.  Direct visualization of colloidal rod assembly by confocal microscopy. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[33]  K. Amako,et al.  Flagellation of Pseudomonas aeruginosa during the Cell Division Cycle , 1982, Microbiology and immunology.

[34]  N. Verstraeten,et al.  Living on a surface: swarming and biofilm formation. , 2008, Trends in microbiology.

[35]  G. O’Toole,et al.  Innate and induced resistance mechanisms of bacterial biofilms. , 2008, Current Topics in Microbiology and Immunology.

[36]  Paul Stoodley,et al.  Evolving concepts in biofilm infections , 2009, Cellular microbiology.

[37]  R. Harshey,et al.  Bacterial motility on a surface: many ways to a common goal. , 2003, Annual review of microbiology.

[38]  Howard C. Berg,et al.  Direct observation of extension and retraction of type IV pili , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[39]  J. Mattick Type IV pili and twitching motility. , 2002, Annual review of microbiology.

[40]  D. Bhaya,et al.  Type IV pilus biogenesis and motility in the cyanobacterium Synechocystis sp. PCC6803 , 2000, Molecular microbiology.

[41]  O. Nybroe,et al.  A panel of Tn7-based vectors for insertion of the gfp marker gene or for delivery of cloned DNA into Gram-negative bacteria at a neutral chromosomal site. , 2001, Journal of microbiological methods.

[42]  E. Greenberg,et al.  A component of innate immunity prevents bacterial biofilm development , 2002, Nature.

[43]  W. Shi,et al.  Type IV pilus of Myxococcus xanthus is a motility apparatus controlled by the frz chemosensory system , 2000, Current Biology.

[44]  R. Kolter,et al.  Biofilm formation as microbial development. , 2000, Annual review of microbiology.

[45]  S. Molin,et al.  Development and Dynamics of Pseudomonassp. Biofilms , 2000, Journal of bacteriology.

[46]  Samuel S. Wu,et al.  Genetic and functional evidence that Type IV pili are required for social gliding motility in Myxococcus xanthus , 1995, Molecular microbiology.

[47]  Vernita Gordon,et al.  Bacteria Use Type IV Pili to Walk Upright and Detach from Surfaces , 2010, Science.

[48]  Marcus L. Roper,et al.  Bacillus subtilis spreads by surfing on waves of surfactant , 2009, Proceedings of the National Academy of Sciences.

[49]  B. Maier,et al.  Dynamics of type IV pili is controlled by switching between multiple states. , 2009, Biophysical journal.

[50]  C. van Delden,et al.  Swarming of Pseudomonas aeruginosa Is Dependent on Cell-to-Cell Signaling and Requires Flagella and Pili , 2000, Journal of bacteriology.

[51]  J. Costerton,et al.  Bacterial biofilms: a common cause of persistent infections. , 1999, Science.

[52]  Jay X. Tang,et al.  Amplified effect of Brownian motion in bacterial near-surface swimming , 2008, Proceedings of the National Academy of Sciences.

[53]  Michael P. Sheetz,et al.  Pilus retraction powers bacterial twitching motility , 2000, Nature.

[54]  H. Berg The rotary motor of bacterial flagella. , 2003, Annual review of biochemistry.

[55]  A. Merz,et al.  Interactions of pathogenic neisseriae with epithelial cell membranes. , 2000, Annual review of cell and developmental biology.