SFB 766: The Bacterial Cell Envelope: Structure, Function and Infection Interface
Biology
Chemistry
Final Report Abstract
The bacterial envelope determines the shape, surface properties, and solute permeability of bacteria. More- over, it serves as a barrier by which bacteria interact and communicate with each other and the environment thereby exerting a decisive function in bacterial physiology, morphogenesis, transport, and sensitivity or re- sistance towards antimicrobial agents as well as in biotechnological processes. In pathogens, the bacterial surface plays a crucial role in infections and elicits immune reactions. The cell envelope consists of the cyto- plasmic membrane, the cell wall with its appendages, and, in case of Gram-negative bacteria, the outer membrane. Due to its chemical and structural complexity and diversity, insight into the molecular details of bacterial envelopes was rather limited when the CRC766 started in 2007. The biosynthesis of the three- dimensional network of the cell wall, its highly dynamic properties, and its role in both bacterial physiology and bacteria-host interaction were only poorly understood. Work of the CRC on the bacterial cell envelope was initiated because of two reasons. On one hand Tübingen had a long-standing tradition in research into various aspects of the bacterial cell envelope. Besides others Wolfhard Weidel, Joachim Höltje and Volkmar Braun had performed pioneering work on the structure and function of the bacterial envelope. However, research in this field has come to a certain stand still thereafter due to the lack of suitable technologies for investigating the cell envelope in more depth. But, on the other hand, the new –omics technologies, improved analytical methods, progress in structural biology and bioin- formatics offered a great chance to continue with cell envelope research at a new level. Research in this CRC was dedicated to expanding our understanding of the structure, function, and biosyn- thesis of the bacterial cell envelope in an interdisciplinary approach. Furthermore, interactions of the cell envelope with the environment and with the host, its contribution to pathogenicity, and its role in the mode of action of new antibiotics as well as in antibiotic resistance were investigated. These aspects were studied in a broad range of bacteria covering major model organisms as well as important human pathogens. The CRC was structured in two sections, Section A was focused on the synthesis, turnover and function of components of the bacterial cell envelope: by analysing cell wall synthesis and structure we could show among others that cell wall turnover is a permanent, dynamic process which can be targeted by antibiotics, that nanopores in septa of filamentous bacteria are required for cell-cell communication, and are equipped with gates which control closing and opening these pores. Research on antibiotic producing bacteria made it possible to characterize and optimize antibiotics by analysing their biosynthesis and their interaction with cell wall associated enzymes. Furthermore, we could show that gene transfer across streptomycete cell walls is mediated by a process resembling chromosome segregation enabling the horizontal transfer of large DNA fragments; a process which is highly relevant for the evolution of antibiotic biosynthesis gene cluster and associated enzymes. Pioneering research revealed that not only the well-known antibiotic producers such as actinobacteria are able to synthesize natural products which can inhibit pathogens but that also members of the human microbiome have this capacity. Staphylococcus lugdunensis living in the human nose was shown to produce lugdunin, a peptide antibiotic inhibiting the growth of S. aureus. Section B was dealing with all aspects of the interaction of bacteria with host cell via the cell envelope. Stud- ies on new structural data of adhesins, on the assembly of β-barrel proteins in the bacterial cell envelope and in mitochondria, and the immune reaction of plants confronted with bacterial pathogen resulted in a better understanding how bacteria infect their hosts. Research on the bacterial cell wall also gains new importance because of the increasing threat caused by antibiotic-resistant pathogens. The bacterial cell wall is the preferred target for the most effective antibiotics. Research on their mode of action and the resistance mechanisms combined with work on cell wall synthesis provides information, which helps to find new and to improve established antibiotics. In this context, we could identify new resistance mechanisms against lipid II binders, and characterize essential processes in cell envelope synthesis that may constitute novel targets for new antibiotics. The activities of the CRC 766 resulted also in important structural developments: it initiated the foundation of the Interfaculty Institute for Microbiology and Infection Medicine (IMIT) of the University. The successful re- search in the CRC 766 was decisive for the selection of Tübingen as a central partner site of the German Center for Infection Research (DZIF). These achievements finally made it possible to succeed with the pro- posal on “Controlling Microbes to fight Infection” initiative in the excellence competition.
Publications
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Schlag, M, Biswas R, Krismer B, Köhler T, Zoll S, Yu W, Schwarz H, Peschel A, Götz F
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Alt S, Burkard N, Kulik A, Grond S, Heide L
(See online at https://doi.org/10.1016/j.chembiol.2010.12.016) - (2011) Conjugal plasmid transfer in Streptomyces resembles bacterial chromosome segregation by FtsK/SpoIIIE. EMBO J. 30: 2246-2254
Vogelmann J, Ammelburg M, Finger C, Guezguez J, Linke D, Flotenmeyer M, Stierhof YD, Wohlleben W, Muth G
(See online at https://doi.org/10.1038/emboj.2011.121) - (2011) Proteins encoded by the mre gene cluster in Streptomyces coelicolor A3(2) co-operate in spore wall synthesis. Mol. Microbiol. 79:1367-1379
Kleinschnitz E-M, Heichlinger A, Schirner K, Winkler J, Latus A, Maldener I, Wohlleben W, and Muth G
(See online at https://doi.org/10.1111/j.1365-2958.2010.07529.x) - (2012) Complete fibre structures of complex trimeric autotransporter adhesins conserved in enterobacteria. Proc. Natl. Acad. Sci. USA 109: 20907-20912
Hartmann MD, Grin I, Dunin-Horkawicz S, Deiss S, Linke D, Lupas AN, and Hernandez Alvarez B
(See online at https://doi.org/10.1073/pnas.1211872110) - (2013) A cell wall recycling shortcut that bypasses peptidoglycan de novo biosynthesis. Nature Chem. Biol. 9: 491-493
Gisin J, Schneider A, Nägele B, Borisova M, Mayer C
(See online at https://doi.org/10.1038/nchembio.1289) - (2013) A two-step sulfation in antibiotic biosynthesis requires a type III polyketide synthase. Nature Chem. Biol. 9: 610-5
Tang X, Eitel K, Kaysser L, Kulik A, Grond S, Gust B
(See online at https://doi.org/10.1038/nchembio.1310) - (2013) Prokaryotic multicellularity: a nanopore array for bacterial cell communication. FASEB J. 27: 2293-300
Lehner J, Berendt S, Dörsam B, Pérez R, Forchhammer K, Maldener I
(See online at https://doi.org/10.1096/fj.12-225854) - (2013) The Receptor-Like Protein ReMAX of Arabidopsis Detects the Microbe-Associated Molecular Pattern eMax from Xanthomonas. Plant Cell 25: 2330-2340
Jehle AK, Lipschis M, Albert M, Fallahzadeh-Mamaghani V, Fürst U, Mueller K, Felix G
(See online at https://doi.org/10.1105/tpc.113.110833) - (2014) Evolutionary Conservation in Biogenesis of β-Barrel Proteins Allows Mitochondria to Assemble a Functional Bacterial Trimeric Autotransporter Protein. J. Biol. Chem. 289: 29457-29470
Ulrich T, Oberhettinger P, Schütz M, Holzer K, Ramms AS, Linke D, Autenrieth IB, Rapaport D
(See online at https://doi.org/10.1074/jbc.m114.565655) - (2014) Host-induced bacterial cell wall decomposition mediates pattern-triggered immunity in Arabidopsis. elife 3: e01990
Liu X, Grabherr HM, Willmann R, Kolb D, Brunner F, Bertsche U, Kuhner D, Franz-Wachtel M, Amin B, Felix G, Ongena M, Nürnberger T, Gust AA
(See online at https://doi.org/10.7554/elife.01990) - (2014) MapZ marks the division sites and positions FtsZ rings in Streptococcus pneumaniae. Nature 516: 259-62
Fleurie A, Lesterlin C, Manuse S, Zhao C, Cluzel C, Lavergne J-P, Franz-Wachtel M, Macek B, Combet C, Kuru E, Van Nieuwenhze MS, Brun YV, Sherratt D, Grangeasse C
(See online at https://doi.org/10.1038/nature13966) - (2015) The inverse autotransporter intimin exports its passenger domain via a hairpin intermediate. J Biol Chem 290: 1837-49
Oberhettinger P, Leo JC, Linke D, Autenrieth IB, Schütz MS
(See online at https://doi.org/10.1074/jbc.m114.604769) - (2015) The lipid-modifying multiple peptide resistance factor is an oligomer consisting of distinct interacting synthase and flippase subunits. mBio. 6 pii: e02340-14
Ernst CM, Kuhn S, Slavetinsky CJ, Krismer B, Heilbronner S, Gekeler C, Kraus D, Wagner S, Peschel A
(See online at https://doi.org/10.1128/mbio.02340-14) - (2016) Distinct mechanisms contribute to immunity in the lantibiotic NAI-107 producer strain Microbispora ATCC PTA-5024. Environ. Microbiol. 18: 118-132
Pozzi R, Coles M, Linke D, Kulik A, Nega M, Wohlleben W, Stegmann E
(See online at https://doi.org/10.1111/1462-2920.12892) - (2016) Fluorescence microscopy of Streptomyces conjugation suggests DNA-transfer at the lateral walls and reveals the spreading of the plasmid in the recipient mycelium. Environ. Microbiol. 18: 598-608
Thoma L, Vollmer B, Muth G
(See online at https://doi.org/10.1111/1462-2920.13027) - (2016) Human commensals producing a novel antibiotic impair pathogen colonization. Nature 535: 511–516
Zipperer A, Konnerth MC, Laux C, Berscheid A, Janek D, Weidenmaier C, Burian M, Schilling NA, Slaventinsky C, Marschal M, Willmann M, Kalbacher H, Schittek B, Brötz-Oesterhelt H, Grond S, Peschel A, Krismer B
(See online at https://doi.org/10.1038/nature18634) - (2016) Mitochondrial-bacterial hybrids of BamA/Tob55 suggest variable requirements for the membrane integration of β-barrel proteins. Sci. Reports 6: 39053
Pfitzner AK, Steblau N, Ulrich T, Oberhettinger P, Autenrieth IB, Schütz M, Rapaport D
(See online at https://doi.org/10.1038/srep39053) - (2016) Polydiglycosylphosphate Transferase PdtA (SCO2578) of Streptomyces coelicolor A3(2) Is Crucial for Proper Sporulation and Apical Tip Extension under Stress Conditions. Appl. Environ. Microbiol. 82: 5661-72
Sigle S, Steblau N, Wohlleben W, Muth G
(See online at https://doi.org/10.1128/aem.01425-16) - (2016) Regulation by the quorum sensor from Vibrio indicates a receptor function for the membrane anchors of adenylate cyclases. eLife 5:13098
Beltz S, Bassler J, Schultz JE
(See online at https://doi.org/10.7554/elife.13098) - (2016). Alpha/beta coiled coils. eLife 5: e11861
Hartmann MD, Mendler CT, Bassler J, Karamichali I, Ridderbusch O, Lupas AN, Hernandez Alvarez B
(See online at https://doi.org/10.7554/elife.11861) - (2016). Structural and functional characterization of the bacterial type III secretion export apparatus. PLoS Pathog. 12:e1006071
Dietsche T, Tesfazgi Mebrhatu M, Brunner MJ, Abrusci P, Yan J, Franz-Wachtel M, Schärfe C, Zilkenat S, Grin I, Galán JE, Kohlbacher O, Lea S, Macek B, Marlovits TC, Robinson C, Wagner S
(See online at https://doi.org/10.1371/journal.ppat.1006071) - (2017) Characterization of a novel signal transducer element intrinsic to class IIIa/b adenylate cyclases and guanylate cyclases FEBS J. 284: 1204-1217
Ziegler M, Bassler, J., Beltz S, Lupas AN, Schultz JE
(See online at https://doi.org/10.1111/febs.14047) - (2017) Identifying components required for OMP biogenesis as novel targets for anti-infective drugs. Virulence 24:1-20
Weirich J, Bräutigam C, Mühlenkamp M, Franz-Wachtel M, Macek B, Meuskens I, Skurnik M, Leskinen K, Bohn E, Autenrieth I, Schütz M
(See online at https://doi.org/10.1080/21505594.2016.1278333) - (2017) Nonclassical Protein Excretion Is Boosted by PSM alpha-Induced Cell Leakage. Cell Rep. 20: 1278-1286
Ebner P, Luqman A, Reichert S, Hauf K, Popella P, Forchhammer K, Otto M, Götz F
(See online at https://doi.org/10.1016/j.celrep.2017.07.045) - (2017) The N-acetylmuramic acid 6-phosphate phosphatase MupP completes the Pseudomonas peptidoglycan recycling pathway leading to intrinsic fosfomycin resistance. mBio. 8: e00092-17
Borisova M, Gisin J, Mayer C
(See online at https://doi.org/10.1128/mbio.00092-17) - (2017). Wall teichoic acids mediate increased virulence in Staphylococcus aureus. Nature Microbiol. 2:16257
Wanner S, Schade J, Keinhorster D, Weller N, George SE, Kull L, Bauer J, Grau T, Winstel V, Stoy H, Kretschmer D, Kolata J, Wolz C, Broker BM, Weidenmaier C
(See online at https://doi.org/10.1038/nmicrobiol.2016.257) - (2015): Novel anti-infective compound. EP 3072 899 B1, Grant 2018-05-02, US 2018 0155397 A1, WO 2016 151005 A1, ES2682595T3
Krismer B, Peschel A, Grond S, Zipperer A, Konnerth M, Janek D
- (2018) A highly asynchronous developmental program triggered during germination of dormant akinetes of the filamentous diazotrophic cyanobacteria. FEMS Microbiol. Ecol. 94: 1
Perez R, Wörmer LP, Sass P, Maldener, I
(See online at https://doi.org/10.1093/femsec/fix131) - (2018) Gain-of-Function Mutations in the Phospholipid Flippase MprF Confer Specific Daptomycin Resistance. mBio 9. pii: e01659-1
Ernst CM, Slavetinsky CJ, Kuhn S, Hauser JN, Nega M, Mishra NN, Gekeler C, Bayer AS, Peschel A
(See online at https://doi.org/10.1128/mbio.01659-18) - (2018) Methicillin-resistant Staphylococcus aureus alters cell wall glycosylation to evade immunity. Nature 563: 705-709
Gerlach D, Guo Y, De Castro C, Kim SH, Schlatterer K, Xu FF, Pereira C, Seeberger PH, Ali S, Codée J, Sirisarn W, Schulte B, Wolz C, Larsen J, Molinaro A, Lee BL, Xia G, Stehle T, Peschel A
(See online at https://doi.org/10.1038/s41586-018-0730-x) - (2018) Structural basis of cell wall peptidoglycan amidation by the GatD/MurT complex of Staphylococcus aureus. Sci. Rep. 8:12953
Nöldeke ER, Muckenfuss LM, Niemann V, Müller A, Störk E, Zocher G, Schneider T, Stehle T
(See online at https://doi.org/10.1038/s41598-018-31098-x) - (2018) Structure of the core of the type three secretion system export apparatus. Nat. Struct. Mol. Biol. 25: 583-90
Kuhlen L, Abrusci P, Johnson S, Gault J, Deme J, Caesar J, Dietsche T, Tesfazgi Mebrhatu M., Ganief T, Macek B, Wagner S, Robinson CV, Lea SM
(See online at https://doi.org/10.1038/s41594-018-0086-9) - (2013): [S,S]-EDDS Biosynthesegene und –proteine und Verfahren zur Biosynthese von [S,S]-EDDS. 14765893.4 (erteilt 6.6.19)
Stegmann E, Wohlleben W, Spohn M, Weber
- (2019) Deprivation of the Periplasmic Chaperone SurA Reduces Virulence and Restores Antibiotic Susceptibility of Multidrug-Resistant Pseudomonas aeruginosa. Front. Microbiol. 10: 100
Klein K, Sonnabend MS, Frank L, Leibiger K, Franz-Wachtel M, Macek B, Trunk T, Leo JC, Autenrieth IB, Schütz M, Bohn E
(See online at https://doi.org/10.3389/fmicb.2019.00100) - (2019) Environmental and cellular factors affecting the localization of T6SS proteins in Burkholderia thailandensis. Int. J. Med. Microbiol. 18: 151335
Lennings J, Makhlouf M, Olejnik P, Mayer C, Brötz-Oesterhelt H, Schwarz S
(See online at https://doi.org/10.1016/j.ijmm.2019.151335) - (2019) Kistamicin biosynthesis reveals the biosynthetic requirements for production of highly crosslinked glycopeptide antibiotics. Nature Commun. 10: 2613
Greule A, Izoré T, Iftime D, Tailhades J, Schoppet M, Zhao Y, Peschke M, Ahmed I, Kulik A, Adamek M, Goode RJA, Schittenhelm RB, Kaczmarski JA, Jackson CJ, Ziemert N, Krenske EH, De Voss JJ, Stegmann E, Cryle MJ
(See online at https://doi.org/10.1038/s41467-019-10384-w) - (2019) Revisiting the regulation of the capsular polysaccharide biosynthesis gene cluster in Staphylococcus aureus. Mol. Microbiol.
Keinhörster D, Salzer A, Duque-Jaramillo A, George SE, Marincola G, Lee JC, Weidenmaier C, Wolz C
(See online at https://doi.org/10.1111/mmi.14347) - (2019) Structure and Function of a Bacterial Gap Junction Analog. Cell 178: 374-384
Weiss GL, Kieninger A-K, Maldener I, Forchhammer K., Pilhofer M
(See online at https://doi.org/10.1016/j.cell.2019.05.055) - (2019) The Burkholderia Type VI Secretion System 5: Composition, Regulation and Role in Virulence. Front. Microbiol. 9:3339
Lennings J, West TE, Schwarz S
(See online at https://doi.org/10.3389/fmicb.2018.03339) - (2019) Two DevBCA-like ABC transporters are involved in the multidrug resistance of the cyanobacterium Anabaena sp. PCC 7120. FEBS Lett. 593: 1818-1826
Shvarev D, Nishi CN, Maldener I
(See online at https://doi.org/10.1002/1873-3468.13450) - (2019). Synthetic lugdunin analogues reveal essential structural motifs for antimicrobial action and proton translocation capability. Angew. Chem. Int. Ed. Engl. 58: 9234-9238
Schilling NA, Berscheid A, Schumacher J, Saur JS, Konnerth MC, Wirtz SN, Beltrán Beleña JM, Zipperer A, Krismer B, Peschel A, Kalbacher H, Brötz-Oesterhelt H, Steinem C, Grond SC
(See online at https://doi.org/10.1002/anie.201901589)
