Near Surface Swimming of Salmonella Typhimurium Explains Target-Site Selection and Cooperative Invasion

Targeting of permissive entry sites is crucial for bacterial infection. The targeting mechanisms are incompletely understood. We have analyzed target-site selection by S. Typhimurium. This enteropathogenic bacterium employs adhesins (e.g. fim) and the type III secretion system 1 (TTSS-1) for host cell binding, the triggering of ruffles and invasion. Typically, S. Typhimurium invasion is focused on a subset of cells and multiple bacteria invade via the same ruffle. It has remained unclear how this is achieved. We have studied target-site selection in tissue culture by time lapse microscopy, movement pattern analysis and modeling. Flagellar motility (but not chemotaxis) was required for reaching the host cell surface in vitro. Subsequently, physical forces trapped the pathogen for ∼1.5–3 s in “near surface swimming”. This increased the local pathogen density and facilitated “scanning” of the host surface topology. We observed transient TTSS-1 and fim-independent “stopping” and irreversible TTSS-1-mediated docking, in particular at sites of prominent topology, i.e. the base of rounded-up cells and membrane ruffles. Our data indicate that target site selection and the cooperative infection of membrane ruffles are attributable to near surface swimming. This mechanism might be of general importance for understanding infection by flagellated bacteria.

[1]  B. Stecher,et al.  Flagella and Chemotaxis Are Required for Efficient Induction of Salmonella enterica Serovar Typhimurium Colitis in Streptomycin-Pretreated Mice , 2004, Infection and Immunity.

[2]  K. Darwin,et al.  Molecular Basis of the Interaction ofSalmonella with the Intestinal Mucosa , 1999, Clinical Microbiology Reviews.

[3]  Piotr Garstecki,et al.  Escherichia coli swim on the right-hand side , 2005, Nature.

[4]  Malte Hermansson,et al.  The DLVO theory in microbial adhesion , 1999 .

[5]  T. Wallis,et al.  Absence of All Components of the Flagellar Export and Synthesis Machinery Differentially Alters Virulence of Salmonella enterica Serovar Typhimurium in Models of Typhoid Fever, Survival in Macrophages, Tissue Culture Invasiveness, and Calf Enterocolitis , 2001, Infection and Immunity.

[6]  M. Woodward,et al.  Membrane ruffling and invasion of human and avian cell lines is reduced for aflagellate mutants of Salmonella enterica serotype Enteritidis. , 2003, International journal of medical microbiology : IJMM.

[7]  C. Ginocchio,et al.  Identification and molecular characterization of a Salmonella typhimurium gene involved in triggering the internalization of salmonellae into cultured epithelial cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[8]  E. Allen-Vercoe,et al.  Fimbriae- and flagella-mediated association with and invasion of cultured epithelial cells by Salmonella enteritidis. , 1999, Microbiology.

[9]  B. Wanner,et al.  One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. Cookson,et al.  In vivo, fliC expression by Salmonella enterica serovar Typhimurium is heterogeneous, regulated by ClpX, and anatomically restricted , 2006, Molecular microbiology.

[11]  S. Banwart,et al.  The polymer physics and chemistry of microbial cell attachment and adhesion. , 2008, Faraday discussions.

[12]  Wolf-Dietrich Hardt,et al.  Role of the Salmonella Pathogenicity Island 1 (SPI-1) Protein InvB in Type III Secretion of SopE and SopE2, Two Salmonella Effector Proteins Encoded Outside of SPI-1 , 2003, Journal of bacteriology.

[13]  S. Kudo,et al.  Bacterial swimming speed and rotation rate of bundled flagella. , 2001, FEMS microbiology letters.

[14]  M. Heikenwalder,et al.  The S. Typhimurium effector SopE induces caspase-1 activation in stromal cells to initiate gut inflammation. , 2009, Cell host & microbe.

[15]  E. McGhie,et al.  Salmonella takes control: effector-driven manipulation of the host , 2009, Current opinion in microbiology.

[16]  A. Takeuchi.  Electron microscope studies of experimental Salmonella infection. I. Penetration into the intestinal epithelium by Salmonella typhimurium. , 1967, The American journal of pathology.

[17]  L. Hernandez,et al.  A Salmonella inositol polyphosphatase acts in conjunction with other bacterial effectors to promote host cell actin cytoskeleton rearrangements and bacterial internalization , 2001, Molecular microbiology.

[18]  Judith P. Armitage,et al.  Signal processing in complex chemotaxis pathways , 2011, Nature Reviews Microbiology.

[19]  Christian von Mering,et al.  Rnai Screen of Salmonella Invasion Shows Role of Copi in Membrane Targeting of Cholesterol and Cdc42 , 2022 .

[20]  Wolf-Dietrich Hardt,et al.  Gut inflammation can boost horizontal gene transfer between pathogenic and commensal Enterobacteriaceae , 2012, Proceedings of the National Academy of Sciences.

[21]  Christine Josenhans,et al.  The role of motility as a virulence factor in bacteria. , 2002, International journal of medical microbiology : IJMM.

[22]  Anne E Carpenter,et al.  CellProfiler: image analysis software for identifying and quantifying cell phenotypes , 2006, Genome Biology.

[23]  K. Jarrell,et al.  The surprisingly diverse ways that prokaryotes move , 2008, Nature Reviews Microbiology.

[24]  Péter Horváth,et al.  Enhanced CellClassifier: a multi-class classification tool for microscopy images , 2010, BMC Bioinformatics.

[25]  Wolf-Dietrich Hardt,et al.  Salmonella type III secretion effectors: pulling the host cell's strings. , 2006, Current opinion in microbiology.

[26]  M. Heikenwalder,et al.  Salmonella Transiently Reside in Luminal Neutrophils in the Inflamed Gut , 2012, PloS one.

[27]  A. Bäumler,et al.  The attenuated sopB mutant of Salmonella enterica serovar Typhimurium has the same tissue distribution and host chemokine response as the wild type in bovine Peyer's patches. , 2003, Veterinary microbiology.

[28]  J. Galán,et al.  The Salmonella typhimurium invasion genes invF and invG encode homologues of the AraC and PulD family of proteins , 1994, Molecular microbiology.

[29]  Steffen Jung,et al.  Microbe sampling by mucosal dendritic cells is a discrete, MyD88-independent stepin ΔinvG S. Typhimurium colitis , 2008, The Journal of experimental medicine.

[30]  M. Surette,et al.  Signal transduction in bacterial chemotaxis , 1992, The Journal of biological chemistry.

[31]  H. C. van der Mei,et al.  Forces involved in bacterial adhesion to hydrophilic and hydrophobic surfaces. , 2008, Microbiology.

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

[33]  K. Schuebel,et al.  S. typhimurium Encodes an Activator of Rho GTPases that Induces Membrane Ruffling and Nuclear Responses in Host Cells , 1998, Cell.

[34]  George M. Hilliard,et al.  Enterotoxigenic Escherichia coli EtpA mediates adhesion between flagella and host cells , 2008, Nature.

[35]  B. Finlay,et al.  Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. , 1990, The Journal of infectious diseases.

[36]  Gang Chen,et al.  Microbial surface thermodynamics and applications. , 2003, Research in microbiology.

[37]  J. Galán,et al.  The Flagellar Sigma Factor FliA (ς28) Regulates the Expression of Salmonella Genes Associated with the Centisome 63 Type III Secretion System , 2000, Infection and Immunity.

[38]  B. Finlay,et al.  Cytoskeletal rearrangements accompanying salmonella entry into epithelial cells. , 1991, Journal of cell science.

[39]  B. Stecher,et al.  Real-time imaging of type III secretion: Salmonella SipA injection into host cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[40]  S. Falkow,et al.  Invasion by Salmonella typhimurium is affected by the direction of flagellar rotation , 1992, Infection and immunity.

[41]  T. C. Weber,et al.  Salmonella gut invasion involves TTSS-2-dependent epithelial traversal, basolateral exit, and uptake by epithelium-sampling lamina propria phagocytes. , 2012, Cell host & microbe.

[42]  Emma Allen-Vercoe,et al.  Lack of flagella disadvantages Salmonella enterica serovar Enteritidis during the early stages of infection in the rat. , 2003, Journal of medical microbiology.

[43]  Michael Wagner,et al.  Reversible and Irreversible Adhesion of Motile Escherichia coli Cells Analyzed by Total Internal Reflection Aqueous Fluorescence Microscopy , 2002, Applied and Environmental Microbiology.

[44]  Jean-Christophe Olivo-Marin,et al.  Extraction of spots in biological images using multiscale products , 2002, Pattern Recognit..

[45]  S. Normark,et al.  Bacterial adhesins in host-microbe interactions. , 2009, Cell host & microbe.

[46]  J. Galán,et al.  Salmonella enterica Serovar Typhimurium Pathogenicity Island 1-Encoded Type III Secretion System Translocases Mediate Intimate Attachment to Nonphagocytic Cells , 2009, Infection and Immunity.

[47]  P. Jenny,et al.  Stochastic modeling of light scattering with fluorescence using a Monte Carlo-based multiscale approach. , 2009, Journal of the Optical Society of America. A, Optics, image science, and vision.

[48]  P. Ganatos,et al.  The motion of a rigid body in viscous fluid bounded by a plane wall , 1989, Journal of Fluid Mechanics.

[49]  W. Hardt,et al.  A substrate of the centisome 63 type III protein secretion system of Salmonella typhimurium is encoded by a cryptic bacteriophage. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[50]  V. Koronakis,et al.  Deciphering Interplay between Salmonella Invasion Effectors , 2008, PLoS pathogens.

[51]  J. Galán,et al.  Manipulation of the host actin cytoskeleton by Salmonella--all in the name of entry. , 2005, Current opinion in microbiology.

[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]  H. Lockman,et al.  Salmonella typhimurium mutants lacking flagella or motility remain virulent in BALB/c mice , 1990, Infection and immunity.

[54]  S. Akira,et al.  The Salmonella Pathogenicity Island (SPI)-2 and SPI-1 Type III Secretion Systems Allow Salmonella Serovar typhimurium to Trigger Colitis via MyD88-Dependent and MyD88-Independent Mechanisms1 , 2005, The Journal of Immunology.

[55]  Thomas Wild,et al.  Machine Learning Improves the Precision and Robustness of High-Content Screens , 2011, Journal of biomolecular screening.

[56]  Stephen J. Smith,et al.  Ruffles induced by Salmonella and other stimuli direct macropinocytosis of bacteria , 1993, Nature.

[57]  Balamurugan Periaswamy,et al.  Salmonella enterica Serovar Typhimurium Binds to HeLa Cells via Fim-Mediated Reversible Adhesion and Irreversible Type Three Secretion System 1-Mediated Docking , 2010, Infection and Immunity.

[58]  W. Rabsch,et al.  Motility allows S. Typhimurium to benefit from the mucosal defence , 2008, Cellular microbiology.

[59]  B. Finlay,et al.  Penetration of Salmonella through a polarized Madin-Darby canine kidney epithelial cell monolayer , 1988, The Journal of cell biology.

[60]  B. Stocker,et al.  Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines , 1981, Nature.

[61]  W. Hardt,et al.  In Macrophages, Caspase-1 Activation by SopE and the Type III Secretion System-1 of S. Typhimurium Can Proceed in the Absence of Flagellin , 2010, PloS one.

[62]  H. Schmieger.  Phage P22-mutants with increased or decreased transduction abilities , 2004, Molecular and General Genetics MGG.

[63]  B. Stocker,et al.  Roles of motility, chemotaxis, and penetration through and growth in intestinal mucus in the ability of an avirulent strain of Salmonella typhimurium to colonize the large intestine of streptomycin-treated mice , 1988, Infection and immunity.