Control of bacterial morphology and differentiation

Summary

Principal Investigator: Yves V Brun
Abstract: DESCRIPTION (provided by applicant): Bacteria undergo specific cell shape changes and localize various proteins to subcellular sites, both of which are important for their survival in te environment. For example, filamentation of uropathogenic E. coli subverts innate defenses during the infection process, and the helical shape of Helicobacter pylori is important for colonization of the stomach. Understanding how bacteria achieve changes in morphology and modes of growth and couple them to the localization of proteins, such as virulence factors, is critical for our ability to inhibit their persistence, proliferation, and host infection. The generl goal of this research is to study the mechanisms that generate various cell shapes and growth patterns and how they are coordinated with protein localization and function. This study takes advantage of stalk synthesis, a specialized zonal mode of growth in Caulobacter crescentus and its relatives that generates thin extensions of the cell envelope and of the recently discovered reproduction of bacteria in the Rhizobiales by polar growth, including some human pathogens. The project has three major aims. The first aim is to determine the role of the penicillin binding protein PbpC in the recruitment of the StpX protein to the stalk. This will be achieved by determining if StpX is inserted into the stalk concurrently with stalk elongation and if PbpC recruits StpX to the stalk by protein-protein interaction or through its enzymatic activity and modification of stalk peptidoglycan. A novel high throughput microscopy screen will be used to identify genes involved in protein targeting to the stalk and in stalk synthesis. The second aim is to determine how the muramidase SpmX is required for stalk synthesis in Asticcacaulis biprosthecum and directs its subcellular location. This will be accomplished by studying the muramidase activity of SpmX and determining its requirement for stalk synthesis and localization, for its own localization, and for the localization of the developmental regulator Div. Protein chimeras will be constructed to determine how the domains of SpmX have evolved to generate species-specific elements of each function. The effect of altering the localization of developmental regulators on developmental outcomes will also be studied. The third aim is to determine how bacteria in the Rhizobiales, including some human pathogens, reproduce by zonal growth at their pole. This will be accomplished by using Agrobacterium tumefaciens as a model to determine the growth mechanism in this group. Genes required to direct polar growth will be identified by candidate and high throughput approaches. Peptidoglycan composition will be analyzed in wild-type and specific mutants to determine if polar growth requires peptidoglycan synthesis and composition that are different than for lateral cell wall synthesis. Finally, the mechanisms by which peptidoglycan synthesis is redirected during the cell cycle will be determined. Insights gained from these studies can be used to design strategies to inhibit growth, prevent key morphological changes, or alter important protein localization pathways in pathogens, thereby improving our ability to control them.
Funding Period: 1995-01-01 - 2016-02-29
more information: NIH RePORT

Top Publications

  1. ncbi Nuclear phosphoinositide kinases and inositol phospholipids
    Michael L Gonzales
    Department of Pharmacology, Program in Molecular and Cellular Pharmacology, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
    J Cell Biochem 97:252-60. 2006
  2. pmc The scaffolding and signalling functions of a localization factor impact polar development
    Patrick D Curtis
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Mol Microbiol 84:712-35. 2012
  3. pmc Microfluidic device for automated synchronization of bacterial cells
    Seth M Madren
    Department of Chemistry, Indiana University, Bloomington, Indiana 47405 7102, United States
    Anal Chem 84:8571-8. 2012
  4. pmc In Situ probing of newly synthesized peptidoglycan in live bacteria with fluorescent D-amino acids
    Erkin Kuru
    Indiana University, Bloomington, IN 47405, USA
    Angew Chem Int Ed Engl 51:12519-23. 2012
  5. pmc General protein diffusion barriers create compartments within bacterial cells
    Susan Schlimpert
    Max Planck Research Group Prokaryotic Cell Biology, Max Planck Institute for Terrestrial Microbiology, Karl von Frisch Strasse 10, 35043 Marburg, Germany
    Cell 151:1270-82. 2012
  6. pmc Peptidoglycan transformations during Bacillus subtilis sporulation
    Elitza I Tocheva
    Division of Biology, California Institute of Technology, 1200 E California Blvd, Pasadena, CA 91125, USA
    Mol Microbiol 88:673-86. 2013
  7. pmc Bypassing the need for subcellular localization of a polysaccharide export-anchor complex by overexpressing its protein subunits
    JUNE JAVENS
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Mol Microbiol 89:350-71. 2013
  8. pmc Biological consequences and advantages of asymmetric bacterial growth
    David T Kysela
    Department of Biology, Indiana University, Bloomington, Indiana 47405 email
    Annu Rev Microbiol 67:417-35. 2013
  9. pmc Effect of a ctrA promoter mutation, causing a reduction in CtrA abundance, on the cell cycle and development of Caulobacter crescentus
    Patrick D Curtis
    Department of Biology, University of Mississippi, 402 Shoemaker, University, MS 38677, USA
    BMC Microbiol 13:166. 2013
  10. pmc Modes of cell wall growth differentiation in rod-shaped bacteria
    Felipe Cava
    Department of Molecular Biology and Laboratory for Molecular Infection Medicine Sweden, Umeå Centre for Microbial Research, Umea University, Umea, Sweden Electronic address
    Curr Opin Microbiol 16:731-7. 2013

Detail Information

Publications37

  1. ncbi Nuclear phosphoinositide kinases and inositol phospholipids
    Michael L Gonzales
    Department of Pharmacology, Program in Molecular and Cellular Pharmacology, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
    J Cell Biochem 97:252-60. 2006
    ..Here, we will introduce the components of nuclear inositol phospholipid signal transduction and discuss how their spatial arrangement may dictate which nuclear functions they are modulating...
  2. pmc The scaffolding and signalling functions of a localization factor impact polar development
    Patrick D Curtis
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Mol Microbiol 84:712-35. 2012
    ..As a whole, the results show that PodJ's scaffolding role for structural and signalling proteins both contribute to flagellar pole organelle development...
  3. pmc Microfluidic device for automated synchronization of bacterial cells
    Seth M Madren
    Department of Chemistry, Indiana University, Bloomington, Indiana 47405 7102, United States
    Anal Chem 84:8571-8. 2012
    ....
  4. pmc In Situ probing of newly synthesized peptidoglycan in live bacteria with fluorescent D-amino acids
    Erkin Kuru
    Indiana University, Bloomington, IN 47405, USA
    Angew Chem Int Ed Engl 51:12519-23. 2012
    ..HCC = 7-hydroxycoumarin, NBD = 7-nitrobenzofurazan, TAMRA = carboxytetramethylrhodamine...
  5. pmc General protein diffusion barriers create compartments within bacterial cells
    Susan Schlimpert
    Max Planck Research Group Prokaryotic Cell Biology, Max Planck Institute for Terrestrial Microbiology, Karl von Frisch Strasse 10, 35043 Marburg, Germany
    Cell 151:1270-82. 2012
    ..Their presence is critical for cellular fitness because they minimize the effective cell volume, allowing faster adaptation to environmental changes that require de novo synthesis of envelope proteins...
  6. pmc Peptidoglycan transformations during Bacillus subtilis sporulation
    Elitza I Tocheva
    Division of Biology, California Institute of Technology, 1200 E California Blvd, Pasadena, CA 91125, USA
    Mol Microbiol 88:673-86. 2013
    ..Endopeptidase activity may be the main switch that governs whether a thin or a thick PG layer is assembled...
  7. pmc Bypassing the need for subcellular localization of a polysaccharide export-anchor complex by overexpressing its protein subunits
    JUNE JAVENS
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Mol Microbiol 89:350-71. 2013
    ..Our results highlight the fact that multiple facets of subcellular localization can be coupled to improve the phenotypic outcome of a protein assembly. ..
  8. pmc Biological consequences and advantages of asymmetric bacterial growth
    David T Kysela
    Department of Biology, Indiana University, Bloomington, Indiana 47405 email
    Annu Rev Microbiol 67:417-35. 2013
    ..We propose specific experimental approaches to further develop our understanding of the prevalence and the ultimate importance of asymmetric bacterial growth. ..
  9. pmc Effect of a ctrA promoter mutation, causing a reduction in CtrA abundance, on the cell cycle and development of Caulobacter crescentus
    Patrick D Curtis
    Department of Biology, University of Mississippi, 402 Shoemaker, University, MS 38677, USA
    BMC Microbiol 13:166. 2013
    ..However, the mechanism by which these mutations affect polar development is not well understood. The aim of this study was to identify new regulators that control multiple aspects of polar organelle development...
  10. pmc Modes of cell wall growth differentiation in rod-shaped bacteria
    Felipe Cava
    Department of Molecular Biology and Laboratory for Molecular Infection Medicine Sweden, Umeå Centre for Microbial Research, Umea University, Umea, Sweden Electronic address
    Curr Opin Microbiol 16:731-7. 2013
    ..Here we review the latest developments, focusing on the impact of new techniques on the precise mapping of cell wall growth sites. ..
  11. pmc Co-ordinate synthesis and protein localization in a bacterial organelle by the action of a penicillin-binding-protein
    H Velocity Hughes
    Department of Biology, Indiana University, Bloomington, IN, 47405, USA
    Mol Microbiol 90:1162-77. 2013
    ....
  12. pmc Discovery of chlamydial peptidoglycan reveals bacteria with murein sacculi but without FtsZ
    Martin Pilhofer
    1 Division of Biology, California Institute of Technology, Pasadena, California 91125, USA 2 Howard Hughes Medical Institute, Pasadena, California 91125, USA 3
    Nat Commun 4:2856. 2013
    ..They also present a unique example of a bacterium with a PG sacculus but without FtsZ, challenging the current hypothesis that it is the absence of a cell wall that renders FtsZ non-essential. ..
  13. pmc A new metabolic cell-wall labelling method reveals peptidoglycan in Chlamydia trachomatis
    G W Liechti
    1 Department of Microbiology and Immunology, F Edward Hebert School of Medicine, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, Maryland 20814 4799, USA 2
    Nature 506:507-10. 2014
    ..These findings culminate 50 years of speculation and debate concerning the chlamydial anomaly and are the strongest evidence so far that chlamydial species possess functional PG. ..
  14. pmc A versatile class of cell surface directional motors gives rise to gliding motility and sporulation in Myxococcus xanthus
    Morgane Wartel
    Laboratoire de Chimie Bacterienne, CNRS UMR 7283, Aix Marseille Universite, Institut de Microbiologie de la Mediterranee, Marseille, France
    PLoS Biol 11:e1001728. 2013
    ..Thus, the Agl-Glt/Nfs machineries may constitute a novel class of directional bacterial surface transporters that can be diversified to specific tasks depending on the cognate cargo and machinery-specific accessories. ..
  15. pmc GeneclusterViz: a tool for conserved gene cluster visualization, exploration and analysis
    Vikas R Pejaver
    School of Informatics and Computing, Indiana University Bloomington, IN 47404, USA
    Bioinformatics 28:1527-9. 2012
    ..Therefore, a software tool that enables visualization and functional analyses of gene clusters would be a great asset to the biological research community...
  16. pmc Polar growth in the Alphaproteobacterial order Rhizobiales
    Pamela J B Brown
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Proc Natl Acad Sci U S A 109:1697-701. 2012
    ..We find that unipolar growth is an ancestral and conserved trait among the Rhizobiales, which includes important mutualists and pathogens of plants and animals...
  17. pmc Surface contact stimulates the just-in-time deployment of bacterial adhesins
    Guanglai Li
    Physics Department, Brown University, Providence, RI 02912, USA
    Mol Microbiol 83:41-51. 2012
    ..This mechanism restricts stable attachment to intimate surface interactions, thereby maximizing surface attachment, discouraging non-productive self-adherence, and preventing curing of the adhesive...
  18. ncbi Dissection of functional domains of the polar localization factor PodJ in Caulobacter crescentus
    Melanie L Lawler
    Department of Biology, Indiana University, 1001 E Third Street, Bloomington, IN 47405 7005, USA
    Mol Microbiol 59:301-16. 2006
    ..Finally, we identify a cytoplasmic region of PodJ involved in targeting it to the flagellar pole, and a periplasmic region required for localization of PleC...
  19. pmc Mutations in DivL and CckA rescue a divJ null mutant of Caulobacter crescentus by reducing the activity of CtrA
    Deanne L Pierce
    Indiana University, Department of Biology, 1001 E 3rd Street, Bloomington, IN 47405, USA
    J Bacteriol 188:2473-82. 2006
    ..In vivo phosphorylation assays confirmed that divJ mutants have elevated levels of CtrA phosphorylation and that this level is reduced in the suppressors with mutations in divL...
  20. pmc Adhesion of single bacterial cells in the micronewton range
    Peter H Tsang
    Department of Physics and Division of Engineering, Brown University, Providence, RI 02912, USA
    Proc Natl Acad Sci U S A 103:5764-8. 2006
    ..To our knowledge, this strength of adhesion is the strongest ever measured for biological adhesives...
  21. pmc A nutrient uptake role for bacterial cell envelope extensions
    Jennifer K Wagner
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Proc Natl Acad Sci U S A 103:11772-7. 2006
    ..crescentus thrives. This advantage is due to the fact that a stalk is long and thin, a favorable shape for maximizing contact with diffusing nutrients while minimizing increases in both surface area and cell volume...
  22. ncbi Out on a limb: how the Caulobacter stalk can boost the study of bacterial cell shape
    Jennifer K Wagner
    Division of Infectious Diseases, Massachusetts General Hospital Harvard Medical School, Cambridge, MA 02139, USA
    Mol Microbiol 64:28-33. 2007
    ....
  23. pmc The structure of FtsZ filaments in vivo suggests a force-generating role in cell division
    Zhuo Li
    Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA
    EMBO J 26:4694-708. 2007
    ....
  24. pmc Advantages and mechanisms of polarity and cell shape determination in Caulobacter crescentus
    Melanie L Lawler
    Department of Biology, Indiana University, 1001 East Third Street, Bloomington, IN 47405 3700, USA
    Curr Opin Microbiol 10:630-7. 2007
    ..Major progress has been made in understanding the regulatory network that coordinates polar development and morphogenesis and the role of polar localization of regulatory proteins...
  25. pmc Characterization of the Caulobacter crescentus holdfast polysaccharide biosynthesis pathway reveals significant redundancy in the initiating glycosyltransferase and polymerase steps
    Evelyn Toh
    Department of Biology, Indiana University, Bloomington, Indiana 47405 3700, USA
    J Bacteriol 190:7219-31. 2008
    ....
  26. pmc Complex regulatory pathways coordinate cell-cycle progression and development in Caulobacter crescentus
    Pamela J B Brown
    Department of Biology, Indiana University, Bloomington, IN 47405 3700, USA
    Adv Microb Physiol 54:1-101. 2009
    ..We examine the recent advances that have enhanced our understanding of the mechanisms of temporal and spatial regulation that occur during cell-cycle progression...
  27. pmc Getting in the loop: regulation of development in Caulobacter crescentus
    Patrick D Curtis
    Department of Biology, Indiana University, 1001 E 3rd St, Bloomington, IN 47405, USA
    Microbiol Mol Biol Rev 74:13-41. 2010
    ..While much has been discovered about Caulobacter crescentus development, areas of potential future research are also highlighted...
  28. pmc Protein localization and dynamics within a bacterial organelle
    H Velocity Hughes
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    Proc Natl Acad Sci U S A 107:5599-604. 2010
    ..Thus, at the submicron scale the localization and the mobility of a protein are precisely regulated in space and time and are important for the correct organization of a subcellular compartment or organelle such as the stalk...
  29. pmc A localized multimeric anchor attaches the Caulobacter holdfast to the cell pole
    Gail G Hardy
    Department of Biology, Jordan Hall 142, Indiana University, 1001 E 3rd Street, Bloomington, IN 47405, USA
    Mol Microbiol 76:409-27. 2010
    ..We hypothesize that HfaB secretes HfaA and HfaD across the outer membrane, and the three proteins form a complex anchoring the holdfast to the stalk...
  30. pmc Microchannel-nanopore device for bacterial chemotaxis assays
    Michelle L Kovarik
    Department of Chemistry, Indiana University, Bloomington, Indiana 47405, United States
    Anal Chem 82:9357-64. 2010
    ..3 μM to 1.3 M. Additionally, we make an unanticipated observation of increased turn frequency in a chemotaxis-impaired mutant which provides new insight into the chemotaxis pathway in C. crescentus...
  31. pmc Genome sequences of eight morphologically diverse Alphaproteobacteria
    Pamela J B Brown
    Department of Biology, Indiana University, Bloomington, IN 47405, USA
    J Bacteriol 193:4567-8. 2011
    ..Here we announce the genome sequences of eight alphaproteobacteria, including the first genome sequences of species belonging to the genera Asticcacaulis, Hirschia, Hyphomicrobium, and Rhodomicrobium...
  32. pmc Polarity and the diversity of growth mechanisms in bacteria
    Pamela J B Brown
    Department of Biology, Indiana University, Bloomington, IN 47405, United States
    Semin Cell Dev Biol 22:790-8. 2011
    ..We describe polar growth among diverse bacteria and consider the advantages and consequences of this mode of cell elongation...
  33. pmc Sequential evolution of bacterial morphology by co-option of a developmental regulator
    Chao Jiang
    Department of Biology, Indiana University, Bloomington, Indiana 47405, USA
    Nature 506:489-93. 2014
    ..Therefore, similar evolutionary principles of morphological transitions apply to both single-celled prokaryotes and multicellular eukaryotes. ..

Research Grants30

  1. Elucidation of a bacterial cell shape generating program and pathogenic functions
    NINA REDA SALAMA; Fiscal Year: 2013
    ..pylori as current strains display increasing resistance to existing therapies and fits the mission of NIAID to understand and treat infectious diseases. ..
  2. Pacific NorthWest Regional Center of Excellence (PNWRCE)
    Jay A Nelson; Fiscal Year: 2013
    ..pseudomallei host pathogen response during both the septicemic as well as the intracellular phases of the disease. ..
  3. Effect of Helicobacter pylori cell shape on pathogenesis
    LAURA ELIZABETH MARTINEZ; Fiscal Year: 2013
    ..We will also investigate the mechanisms by which PG recycling mutants cause elevated pro-inflammatory cytokine release (aim 3B). ..
  4. Structural Components of the Campylobacter jejuni Polar Flagellar Motor
    David R Hendrixson; Fiscal Year: 2013
    ....
  5. Membrane Remodeling and Protein Trafficking in Bacteria
    David Z Rudner; Fiscal Year: 2013
    ..Understanding the interplay between membrane proteins and the lipid bilayer in which they resides could lead to the discovery of new targets appropriate for antimicrobial therapeutics. ..
  6. CENTER FOR GASTROINTESTINAL BIOLOGY AND DISEASE
    Robert S Sandler; Fiscal Year: 2013
    ..Through all of its activities, the Center improves communication, promotes collaboration, develops careers and generally enriches the intellectual climate for digestive disease research. ..
  7. Bacterial cytoskeletal structures and dynamics at single molecule level
    Ji Yu; Fiscal Year: 2013
    ..The knowledge gained would improve our fundamental understandings concerning the biology of prokaryotic life, including many prokaryotic pathogens that are important to human health. ..
  8. Assembly and Function of the Septal Ring in E. coli
    PIET A DE BOER; Fiscal Year: 2013
    ..A number of SR proteins are, or are actively pursued as, drug targets. This project may lead to the identification of additional promising targets for, and rational designs of, new anti-bacterial drugs. ..
  9. Peptidoglycan Biogenesis in Escherichia Coli
    Thomas G Bernhardt; Fiscal Year: 2013
    ....
  10. FORESPORE ENGULFMENT DURING B SUBTILIS SPORULATION
    KIT J POGLIANO; Fiscal Year: 2013
    ....