Structure and mechanism of the Nap adhesion complex from the human pathogen Mycoplasma genitalium.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
08 06 2020
08 06 2020
Historique:
received:
22
11
2019
accepted:
06
05
2020
entrez:
10
6
2020
pubmed:
10
6
2020
medline:
19
8
2020
Statut:
epublish
Résumé
Mycoplasma genitalium is a human pathogen adhering to host target epithelial cells and causing urethritis, cervicitis and pelvic inflammatory disease. Essential for infectivity is a transmembrane adhesion complex called Nap comprising proteins P110 and P140. Here we report the crystal structure of P140 both alone and in complex with the N-terminal domain of P110. By cryo-electron microscopy (cryo-EM) and tomography (cryo-ET) we find closed and open Nap conformations, determined at 9.8 and 15 Å, respectively. Both crystal structures and the cryo-EM structure are found in a closed conformation, where the sialic acid binding site in P110 is occluded. By contrast, the cryo-ET structure shows an open conformation, where the binding site is accessible. Structural information, in combination with functional studies, suggests a mechanism for attachment and release of M. genitalium to and from the host cell receptor, in which Nap conformations alternate to sustain motility and guarantee infectivity.
Identifiants
pubmed: 32513917
doi: 10.1038/s41467-020-16511-2
pii: 10.1038/s41467-020-16511-2
pmc: PMC7280502
doi:
Substances chimiques
Bacterial Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2877Références
Burgos, R. et al. Mycoplasma genitalium P140 and P110 cytadhesins are reciprocally stabilized and required for cell adhesion and terminal-organelle development. J. Bacteriol. 188, 8627–37 (2006).
doi: 10.1128/JB.00978-06
Mernaugh, G. R., Dallo, S. F., Holt, S. C. & Baseman, J. B. Properties of adhering and nonadhering populations of Mycoplasma genitalium. Clin. Infect. Dis. 17(Suppl 1), S69–78 (1993).
doi: 10.1093/clinids/17.Supplement_1.S69
Scheffer, M. P. et al. Structural characterization of the NAP; the major adhesion complex of the human pathogen Mycoplasma genitalium. Mol. Microbiol. 105, 869–879 (2017).
Nakane, D., Adan-Kubo, J., Kenri, T. & Miyata, M. Isolation and characterization of P1 adhesin, a leg protein of the gliding bacterium Mycoplasma pneumoniae. J. Bacteriol. 193, 715–22 (2011).
doi: 10.1128/JB.00796-10
Garcia-Morales, L., Gonzalez-Gonzalez, L., Querol, E. & Pinol, J. A minimized motile machinery for Mycoplasma genitalium. Mol. Microbiol 100, 125–38 (2016).
doi: 10.1111/mmi.13305
Aparicio, D. et al. Mycoplasma genitalium adhesin P110 binds sialic-acid human receptors. Nat. Commun. 9, 4471 (2018).
doi: 10.1038/s41467-018-06963-y
Ma, L., Jensen, J. S., Mancuso, M., Myers, L. & Martin, D. H. Kinetics of genetic variation of the Mycoplasma genitalium MG192 Gene in experimentally infected chimpanzees. Infect. Immun. 84, 747–53 (2015).
doi: 10.1128/IAI.01162-15
Wood, G. E. et al. Persistence, immune response, and antigenic variation of Mycoplasma genitalium in an experimentally infected pig-tailed macaque (Macaca nemestrina). Infect. Immun. 81, 2938–51 (2013).
doi: 10.1128/IAI.01322-12
Wood, G. E., Patton, D. L., Cummings, P. K., Iverson-Cabral, S. L. & Totten, P. A. Experimental infection of pig-tailed macaques (Macaca nemestrina) with Mycoplasma genitalium. Infect. Immun. 85, e00738-16.(2017).
Cao, B. et al. High prevalence of macrolide resistance in Mycoplasma pneumoniae isolates from adult and adolescent patients with respiratory tract infection in China. Clin. Infect. Dis. 51, 189–194 (2010).
doi: 10.1086/653535
Parrott, G. L., Kinjo, T. & Fujita, J. A compendium for Mycoplasma pneumoniae. Front. Microbiol. 7, 513 (2016).
pubmed: 27148202
pmcid: 4828434
Bradshaw, C. S. et al. Azithromycin failure in Mycoplasma genitalium urethritis. Emerg. Infect. Dis. 12, 1149–52 (2006).
doi: 10.3201/eid1207.051558
Couldwell, D. L. & Lewis, D. A. Mycoplasma genitalium infection: current treatment options, therapeutic failure, and resistance-associated mutations. Infect. Drug Resist. 8, 147–61 (2015).
pubmed: 26060411
pmcid: 4454211
Ofek, I., Hasty, D. L. & Sharon, N. Anti-adhesion therapy of bacterial diseases: prospects and problems. FEMS Immunol. Med. Microbiol. 38, 181–91 (2003).
doi: 10.1016/S0928-8244(03)00228-1
Sharon, N. Carbohydrates as future anti-adhesion drugs for infectious diseases. Biochim. Biophys. Acta 1760, 527–537 (2006).
doi: 10.1016/j.bbagen.2005.12.008
Pettersen, E. F. et al. UCSF chimera—a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–12 (2004).
doi: 10.1002/jcc.20084
Krissinel, E. & Henrick, K. Inference of macromolecular assemblies from crystalline state. J. Mol. Biol. 372, 774–97 (2007).
doi: 10.1016/j.jmb.2007.05.022
Russ, W. P. & Engelman, D. M. The GxxxG motif: a framework for transmembrane helix-helix association. J. Mol. Biol. 296, 911–9 (2000).
doi: 10.1006/jmbi.1999.3489
Seto, S., Kenri, T., Tomiyama, T. & Miyata, M. Involvement of P1 adhesin in gliding motility of Mycoplasma pneumoniae as revealed by the inhibitory effects of antibody under optimized gliding conditions. J. Bacteriol. 185, 1875–7 (2005).
doi: 10.1128/JB.187.5.1875-1877.2005
Berrow, N. S. et al. A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Res. 35, e45 (2007).
doi: 10.1093/nar/gkm047
Winter, G. xia2: an expert system for macromolecular crystallography data reduction. J. Appl. Crystallogr. 43, 186–190 (2010).
doi: 10.1107/S0021889809045701
Kabsch, W. Xds. Acta Crystallogr D Biol. Crystallogr. 66, 125–32 (2010).
doi: 10.1107/S0907444909047337
Evans, P. Scaling and assessment of data quality. Acta Crystallogr. Sect. D 62, 72–82 (2006).
doi: 10.1107/S0907444905036693
CCP4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr D. Biol. Crystallogr 50, 760–763 (1994).
doi: 10.1107/S0907444994003112
McCoy, A. J. Solving structures of protein complexes by molecular replacement with Phaser. Acta Crystallogr. Sect. D Biol. Crystallogr. 63, 32–41 (2007).
doi: 10.1107/S0907444906045975
Cowtan, K. D. & Main, P. Phase combination and cross validation in iterated density-modification calculations. Acta Crystallogr. Sect. D 52, 43–48 (1996).
doi: 10.1107/S090744499500761X
Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol. Crystallogr 60, 2126–32 (2004).
doi: 10.1107/S0907444904019158
Murshudov, G. N. et al. REFMAC5 for the refinement of macromolecular crystal structures. Acta Crystallogr. Sect. D Biol. Crystallogr. 67, 355–367 (2011).
doi: 10.1107/S0907444911001314
Mastronarde, D. N. Automated electron microscope tomography using robust prediction of specimen movements. J. Struct. Biol. 152, 36–51 (2005).
doi: 10.1016/j.jsb.2005.07.007
Kunz, M. & Frangakis, A. S. Three-dimensional CTF correction improves the resolution of electron tomograms. J. Struct. Biol. 197, 114–122 (2017).
doi: 10.1016/j.jsb.2016.06.016
Pruggnaller, S., Mayr, M. & Frangakis, A. S. A visualization and segmentation toolbox for electron microscopy. J. Struct. Biol. 164, 161–5 (2008).
doi: 10.1016/j.jsb.2008.05.003
Zivanov, J. et al. New tools for automated high-resolution cryo-EM structure determination in RELION-3. eLife 7, e42166 (2018).
doi: 10.7554/eLife.42166
Goddard, T. D. et al. UCSF ChimeraX: meeting modern challenges in visualization and analysis. Protein Sci. 27, 14–25 (2018).
doi: 10.1002/pro.3235
Zheng, S. Q. et al. MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy. Nat. Methods 14, 331–332 (2017).
doi: 10.1038/nmeth.4193
Zhang, K. Gctf: real-time CTF determination and correction. J. Struct. Biol. 193, 1–12 (2016).
doi: 10.1016/j.jsb.2015.11.003
Scheres, S. H. A Bayesian view on cryo-EM structure determination. J. Mol. Biol. 415, 406–418 (2012).
doi: 10.1016/j.jmb.2011.11.010
Pich, O. Q., Burgos, R., Planell, R., Querol, E. & Pinol, J. Comparative analysis of antibiotic resistance gene markers in Mycoplasma genitalium: application to studies of the minimal gene complement. Microbiology 152, 519–527 (2006).
doi: 10.1099/mic.0.28287-0
Garcia-Morales, L., Gonzalez-Gonzalez, L., Costa, M., Querol, E. & Pinol, J. Quantitative assessment of mycoplasma hemadsorption activity by flow cytometry. PLoS ONE 9, e87500 (2014).
doi: 10.1371/journal.pone.0087500