Fluorescence and atomic force microscopy imaging of wall teichoic acids in Lactobacillus plantarum.

Although teichoic acids are major constituents of bacterial cell walls, little is known about the relationships between their spatial localization and their functional roles. Here, we used single-molecule atomic force microscopy (AFM) combined with fluorescence microscopy to image the distribution of wall teichoic acids (WTAs) in Lactobacillus plantarum, in relation with their physiological roles. Phenotype analysis of the wild-type strain and of mutant strains deficient for the synthesis of WTAs (ΔtagO) or cell wall polysaccharides (Δcps1-4) revealed that WTAs are required for proper cell elongation and cell division. Nanoscale imaging by AFM showed that strains expressing WTAs have a highly polarized surface morphology, the poles being much smoother than the side walls. AFM and fluorescence imaging with specific lectin probes demonstrated that the polarized surface structure correlates with a heterogeneous distribution of WTAs, the latter being absent from the surface of the poles. These observations indicate that the polarized distribution of WTAs in L. plantarum plays a key role in controlling cell morphogenesis (surface roughness, cell shape, elongation, and division).

[1]  K. Kurokawa,et al.  Pleiotropic Roles of Polyglycerolphosphate Synthase of Lipoteichoic Acid in Growth of Staphylococcus aureus Cells , 2008, Journal of bacteriology.

[2]  P. Hols,et al.  The biosynthesis and functionality of the cell-wall of lactic acid bacteria , 1999, Antonie van Leeuwenhoek.

[3]  Yves F Dufrêne,et al.  Atomic force microscopy and chemical force microscopy of microbial cells , 2008, Nature Protocols.

[4]  E. Vaughan,et al.  Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation. , 2004, FEMS microbiology letters.

[5]  Daniel J Müller,et al.  Force probing surfaces of living cells to molecular resolution. , 2009, Nature chemical biology.

[6]  V. Dupres,et al.  Structure, cell wall elasticity and polysaccharide properties of living yeast cells, as probed by AFM , 2008, Nanotechnology.

[7]  B. Neumeister,et al.  Role of teichoic acids in Staphylococcus aureus nasal colonization, a major risk factor in nosocomial infections , 2004, Nature Medicine.

[8]  Y. Dufrêne,et al.  Application of X-ray photoelectron spectroscopy to microorganisms , 1994 .

[9]  M. Kleerebezem,et al.  Cre-lox-Based System for Multiple Gene Deletions and Selectable-Marker Removal in Lactobacillus plantarum , 2006, Applied and Environmental Microbiology.

[10]  J. Hay,et al.  Teichoic Acids and the Structure of Bacterial Walls , 1961, Nature.

[11]  Y. Dufrêne,et al.  Detection and localization of single molecular recognition events using atomic force microscopy , 2006, Nature Methods.

[12]  S. D. De Keersmaecker,et al.  Detection, localization, and conformational analysis of single polysaccharide molecules on live bacteria. , 2008, ACS nano.

[13]  C. Weidenmaier,et al.  Teichoic acids and related cell-wall glycopolymers in Gram-positive physiology and host interactions , 2008, Nature Reviews Microbiology.

[14]  P. M. Pereira,et al.  Teichoic acids are temporal and spatial regulators of peptidoglycan cross-linking in Staphylococcus aureus , 2010, Proceedings of the National Academy of Sciences.

[15]  J. Höltje,et al.  Growth of the Stress-Bearing and Shape-Maintaining Murein Sacculus of Escherichia coli , 1998, Microbiology and Molecular Biology Reviews.

[16]  Francis C. Neuhaus,et al.  A Continuum of Anionic Charge: Structures and Functions of d-Alanyl-Teichoic Acids in Gram-Positive Bacteria , 2003, Microbiology and Molecular Biology Reviews.

[17]  T. Irisawa,et al.  Comparison of Components and Synthesis Genes of Cell Wall Teichoic Acid among Lactobacillus plantarum Strains , 2010, Bioscience, biotechnology, and biochemistry.

[18]  Y. Dufrêne,et al.  Surface composition, surface properties, and adhesiveness of Azospirillum brasilense—variation during growth , 1996 .

[19]  O. Schneewind,et al.  Synthesis of glycerol phosphate lipoteichoic acid in Staphylococcus aureus , 2007, Proceedings of the National Academy of Sciences.

[20]  A. L. Koch,et al.  Inside-to-outside growth and turnover of the wall of gram-positive rods. , 1985, Journal of theoretical biology.

[21]  Wold,et al.  The normal Lactobacillus flora of healthy human rectal and oral mucosa , 1998, Journal of applied microbiology.

[22]  V. Dupres,et al.  Stretching polysaccharides on live cells using single molecule force spectroscopy , 2009, Nature Protocols.

[23]  S. Okada,et al.  Structures of Two Monomeric Units of Teichoic Acid Prepared from the Cell Wall of Lactobacillus plantarum NRIC 1068 , 2009, Bioscience, biotechnology, and biochemistry.

[24]  W. Norde,et al.  X-ray photoelectron spectroscopy analysis of whole cells and isolated cell walls of gram-positive bacteria: comparison with biochemical analysis , 1997, Journal of bacteriology.

[25]  E. Brown,et al.  Wall Teichoic Acid Polymers Are Dispensable for Cell Viability in Bacillus subtilis , 2006, Journal of bacteriology.

[26]  J. Errington,et al.  Distinct and essential morphogenic functions for wall‐ and lipo‐teichoic acids in Bacillus subtilis , 2009, The EMBO journal.

[27]  J. Sekiguchi,et al.  The major and minor wall teichoic acids prevent the sidewall localization of vegetative dl‐endopeptidase LytF in Bacillus subtilis , 2008, Molecular microbiology.

[28]  M. Kleerebezem,et al.  Complete genome sequence of Lactobacillus plantarum WCFS1 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[29]  S. Heptinstall,et al.  Teichoic Acids and Membrane Function in Bacteria , 1970, Nature.

[30]  Michiel Kleerebezem,et al.  Identification of Genetic Loci in Lactobacillus plantarum That Modulate the Immune Response of Dendritic Cells Using Comparative Genome Hybridization , 2010, PloS one.

[31]  F. Kienberger,et al.  A new, simple method for linking of antibodies to atomic force microscopy tips. , 2007, Bioconjugate chemistry.

[32]  S. Ahrné,et al.  A mannose-specific adherence mechanism in Lactobacillus plantarum conferring binding to the human colonic cell line HT-29 , 1996, Applied and environmental microbiology.

[33]  K. Amako,et al.  Location of peptidoglycan and teichoic acid on the cell wall surface of Staphylococcus aureus as determined by immunoelectron microscopy. , 1992, Journal of electron microscopy.

[34]  Vesa,et al.  Pharmacokinetics of Lactobacillus plantarum NCIMB 8826, Lactobacillus fermentum KLD, and Lactococcus lactis MG 1363 in the human gastrointestinal tract , 2000, Alimentary pharmacology & therapeutics.

[35]  J. Hermoso,et al.  Pneumococcal CbpD is a murein hydrolase that requires a dual cell envelope binding specificity to kill target cells during fratricide , 2010, Molecular microbiology.

[36]  O. Schneewind,et al.  Genes Required for Glycolipid Synthesis and Lipoteichoic Acid Anchoring in Staphylococcus aureus , 2007, Journal of bacteriology.

[37]  M. Kleerebezem,et al.  d-Alanyl Ester Depletion of Teichoic Acids in Lactobacillus plantarum Results in a Major Modification of Lipoteichoic Acid Composition and Cell Wall Perforations at the Septum Mediated by the Acm2 Autolysin , 2006, Journal of bacteriology.

[38]  A. Singh,et al.  Synthetic lethal compound combinations reveal a fundamental connection between wall teichoic acid and peptidoglycan biosyntheses in Staphylococcus aureus. , 2011, ACS chemical biology.

[39]  Wenjun Zhao,et al.  Lesions in Teichoic Acid Biosynthesis in Staphylococcus aureus Lead to a Lethal Gain of Function in the Otherwise Dispensable Pathway , 2006, Journal of bacteriology.

[40]  A. L. Koch,et al.  Insertion and fate of the cell wall in Bacillus subtilis , 1984, Journal of bacteriology.

[41]  A. Mathias,et al.  Cytidine nucleotides. Part I. Isolation from lactobacillus arabinosus , 1954 .

[42]  Yves F. Dufrêne,et al.  Towards nanomicrobiology using atomic force microscopy , 2008, Nature Reviews Microbiology.

[43]  Julie Gold,et al.  Protein Adsorption on Model Surfaces with Controlled Nanotopography and Chemistry , 2002 .

[44]  Guillaume Andre,et al.  Imaging the nanoscale organization of peptidoglycan in living Lactococcus lactis cells , 2010, Nature communications.

[45]  H. Schwarz,et al.  Role of staphylococcal wall teichoic acid in targeting the major autolysin Atl , 2010, Molecular microbiology.