Artículo

Este artículo es de Acceso Abierto y puede ser descargado en su versión final desde nuestro repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

Phosphatidylcholine (PC), a common phospholipid of the eukaryotic cell membrane, is present in the cell envelope of the intracellular pathogen Brucella abortus, the etiological agent of bovine brucellosis. In this pathogen, the biosynthesis of PC proceeds mainly through the phosphatidylcholine synthase pathway; hence, it relies on the presence of choline in the milieu. These observations imply that B. abortus encodes an as-yet-unknown choline uptake system. Taking advantage of the requirement of choline uptake for PC synthesis, we devised a method that allowed us to identify a homologue of ChoX, the high-affinity periplasmic binding protein of the ABC transporter ChoXWV. Disruption of the choX gene completely abrogated PC synthesis at low choline concentrations in the medium, thus indicating that it is a high-affinity transporter needed for PC synthesis via the PC synthase (PCS) pathway. However, the synthesis of PC was restored when the mutant was incubated in media with higher choline concentrations, suggesting the presence of an alternative low-affinity choline uptake activity. By means of a fluorescence-based equilibrium- binding assay and using the kinetics of radiolabeled choline uptake, we show that ChoX binds choline with an extremely high affinity, and we also demonstrate that its activity is inhibited by increasing choline concentrations. Cell infection assays indicate that ChoX activity is required during the first phase of B. abortus intracellular traffic, suggesting that choline concentrations in the early and intermediate Brucella-containing vacuoles are limited. Altogether, these results suggest that choline transport and PC synthesis are strictly regulated in B. abortus. © 2013, American Society for Microbiology.

Registro:

Documento: Artículo
Título:Identification and characterization of a high-affinity choline uptake system of Brucella abortus
Autor:Herrmann, C.K.; Bukata, L.; Melli, L.; Marchesini, M.I.; Caramelo, J.J.; Comerci, D.J.
Filiación:Instituto de Investigaciones Biotecnológicas Dr. Rodolfo A. Ugalde (IIB-INTECH), Universidad Nacional de San Martín, Consejo Nacional de Investigaciones Científicas y Técnicas (UNSAM-CONICET), San Martín, Buenos Aires, Argentina
Instituto de Investigaciones Bioquímicas de Buenos Aires (IIBBA-CONICET), Fundación Instituto Leloir, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Grupo Pecuario, Comisión Nacional de Energía Atómica (CNEA), Buenos Aires, Argentina
Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, United States
Palabras clave:choline; ChoX protein; periplasmic binding protein; phosphatidylcholine; unclassified drug; article; binding affinity; binding assay; Brucella abortus; cell migration; cell vacuole; choline uptake; controlled study; fluorescence; gene disruption; isotope labeling; mutant; nonhuman; phospholipid synthesis; priority journal; protein analysis; Amino Acid Sequence; Animals; ATP-Binding Cassette Transporters; Bacterial Proteins; Biological Transport, Active; Brucella abortus; Cell Membrane; Choline; Female; Gene Expression Regulation, Bacterial; Macrophages; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Open Reading Frames; Phosphatidylcholines
Año:2013
Volumen:195
Número:3
Página de inicio:493
Página de fin:501
DOI: http://dx.doi.org/10.1128/JB.01929-12
Título revista:Journal of Bacteriology
Título revista abreviado:J. Bacteriol.
ISSN:00219193
CODEN:JOBAA
CAS:choline, 123-41-1, 13232-47-8, 1927-06-6, 4858-96-2, 62-49-7, 67-48-1; phosphatidylcholine, 55128-59-1, 8002-43-5; Bacterial Proteins; Choline, 62-49-7; Phosphatidylcholines
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_00219193_v195_n3_p493_Herrmann.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v195_n3_p493_Herrmann

Referencias:

  • Blusztajn, J.K., Choline, a vital amine (1998) Science, 281, pp. 794-795
  • Michel, V., Yuan, Z., Ramsubir, S., Bakovic, M., Choline transport for phospholipid synthesis (2006) Exp. Biol. Med. (Maywood), 231, pp. 490-504
  • Zeisel, S.H., Da Costa, K.A., Franklin, P.D., Alexander, E.A., Lamont, J.T., Sheard, N.F., Beiser, A., Choline, an essential nutrient for humans (1991) FASEB J., 5, pp. 2093-2098
  • Zeisel, S.H., Choline: essential for brain development and function (1997) Adv. Pediatr., 44, pp. 263-295
  • Aktas, M., Wessel, M., Hacker, S., Klusener, S., Gleichenhagen, J., Narberhaus, F., Phosphatidylcholine biosynthesis and its significance in bacteria interacting with eukaryotic cells (2010) Eur. J. Cell Biol., 89, pp. 888-894
  • Sohlenkamp, C., Lopez-Lara, I.M., Geiger, O., Biosynthesis of phosphatidylcholine in bacteria (2003) Prog. Lipid Res., 42, pp. 115-162
  • Aktas, M., Jost, K.A., Fritz, C., Narberhaus, F., Choline uptake in Agrobacterium tumefaciens by the high-affinity ChoXWV transporter (2011) J. Bacteriol., 193, pp. 5119-5129
  • Comerci, D.J., Altabe, S., de Mendoza, D., Ugalde, R.A., Brucella abortus synthesizes phosphatidylcholine from choline provided by the host (2006) J. Bacteriol., 188, pp. 1929-1934
  • Conde-Alvarez, R., Grilló, M.J., Salcedo, S.P., de Miguel, M.J., Fugier, E., Gorvel, J.P., Moriyón, I., Iriarte, M., Synthesis of phosphatidylcholine, a typical eukaryotic phospholipid, is necessary for full virulence of the intracellular bacterial parasite Brucella abortus (2006) Cell. Microbiol., 8, pp. 1322-1335
  • Conover, G.M., Martinez-Morales, F., Heidtman, M.I., Luo, Z.Q., Tang, M., Chen, C., Geiger, O., Isberg, R.R., Phosphatidylcholine synthesis is required for optimal function of Legionella pneumophila virulence determinants (2008) Cell. Microbiol., 10, pp. 514-528
  • Wessel, M., Klüsener, S., Gödeke, J., Fritz, C., Hacker, S., Narberhaus, F., Virulence of Agrobacterium tumefaciens requires phosphatidylcholine in the bacterial membrane (2006) Mol. Microbiol., 62, pp. 906-915
  • Thiele, O.W., Kehr, W., The "free" lipids of Brucella abortus Bang (1969) Concerning the neutral lipids. Eur. J. Biochem., 9, pp. 167-175. , In German
  • Corbel, M.J., Brucellosis: an overview (1997) Emerg. Infect. Dis., 3, pp. 213-221
  • Moreno, E., Moriyon, I., Brucella melitensis: a nasty bug with hidden credentials for virulence (2002) Proc. Natl. Acad. Sci. U. S. A., 99, pp. 1-3
  • Celli, J., de Chastellier, C., Franchini, D.M., Pizarro-Cerda, J., Moreno, E., Gorvel, J.P., Brucella evades macrophage killing via VirB-dependent sustained interactions with the endoplasmic reticulum (2003) J. Exp. Med., 198, pp. 545-556
  • Moriyón, I., López-Goñi, I., Structure and properties of the outer membranes of Brucella abortus and Brucella melitensis (1998) Int. Microbiol., 1, pp. 19-26
  • Sohlenkamp, C., de Rudder, K.E., Rohrs, V.V., Lopez-Lara, I.M., Geiger, O., Cloning and characterization of the gene for phosphatidylcholine synthase (2000) J. Biol. Chem., 275, p. 27500
  • de Rudder, K.E., Sohlenkamp, C., Geiger, O., Plant-exuded choline is used for rhizobial membrane lipid biosynthesis by phosphatidylcholine synthase (1999) J. Biol. Chem., 274, pp. 20011-20016
  • Kent, C., Gee, P., Lee, S.Y., Bian, X., Fenno, J.C., A CDP-choline pathway for phosphatidylcholine biosynthesis in Treponema denticola (2004) Mol. Microbiol., 51, pp. 471-481
  • Lopez-Lara, I.M., Geiger, O., Novel pathway for phosphatidylcholine biosynthesis in bacteria associated with eukaryotes (2001) J. Biotechnol., 91, pp. 211-221
  • Martinez-Morales, F., Schobert, M., Lopez-Lara, I.M., Geiger, O., Pathways for phosphatidylcholine biosynthesis in bacteria (2003) Microbiology, 149, pp. 3461-3471
  • Dupont, L., Garcia, I., Poggi, M.C., Alloing, G., Mandon, K., Le Rudulier, D.D., The Sinorhizobium meliloti ABC transporter Cho is highly specific for choline and expressed in bacteroids from Medicago sativa nodules (2004) J. Bacteriol., 186, pp. 5988-5996
  • Styrvold, O.B., Falkenberg, P., Landfald, B., Eshoo, M.W., Bjornsen, T., Strom, A.R., Selection, mapping, and characterization of osmoregulatory mutants of Escherichia coli blocked in the choline-glycine betaine pathway (1986) J. Bacteriol., 165, pp. 856-863
  • Chen, C., Malek, A.A., Wargo, M.J., Hogan, D.A., Beattie, G.A., The ATP-binding cassette transporter Cbc (choline/betaine/carnitine) recruits multiple substrate-binding proteins with strong specificity for distinct quaternary ammonium compounds (2010) Mol. Microbiol., 75, pp. 29-45
  • Oswald, C., Smits, S.H., Höing, M., Bremer, E., Schmitt, L., Structural analysis of the choline-binding protein ChoX in a semi-closed and ligandfree conformation (2009) Biol. Chem., 390, pp. 1163-1170
  • Oswald, C., Smits, S.H., Höing, M., Sohn-Bösser, L., Dupont, L., Le Rudulier, D., Schmitt, L., Bremer, E., Crystal structures of the choline/ acetylcholine substrate-binding protein ChoX from Sinorhizobium meliloti in the liganded and unliganded-closed states (2008) J. Biol. Chem., 283, pp. 32848-32859
  • Gerhardt, P., The nutrition of brucellae (1958) Bacteriol. Rev., 22, pp. 81-98
  • Bligh, E.G., Dyer, W.J., A rapid method of total lipid extraction and purification (1959) Can. J. Biochem. Physiol., 37, pp. 911-917
  • Punta, M., Coggill, P.C., Eberhardt, R.Y., Mistry, J., Tate, J., Boursnell, C., Pang, N., Finn, R.D., The Pfam protein families database (2012) Nucleic Acids Res., 40, pp. D290-D301
  • Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., Lipman, D.J., Gapped BLAST and PSI-BLAST: a new generation of protein database search programs (1997) Nucleic Acids Res., 25, pp. 3389-3402
  • Thompson, J.D., Higgins, D.G., Gibson, T.J., CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice (1994) Nucleic Acids Res., 22, pp. 4673-4680
  • Sieira, R., Comerci, D.J., Pietrasanta, L.I., Ugalde, R.A., Integration host factor is involved in transcriptional regulation of the Brucella abortus virB operon (2004) Mol. Microbiol., 54, pp. 808-822
  • Marchesini, M.I., Ugalde, J.E., Czibener, C., Comerci, D.J., Ugalde, R.A., N-terminal-capturing screening system for the isolation of Brucella abortus genes encoding surface exposed and secreted proteins (2004) Microb. Pathog., 37, pp. 95-105
  • Celli, J., Salcedo, S.P., Gorvel, J.P., Brucella coopts the small GTPase Sar1 for intracellular replication (2005) Proc. Natl. Acad. Sci. U. S. A., 102, pp. 1673-1678
  • Kohler, S., Foulongne, V., Ouahrani-Bettache, S., Bourg, G., Teyssier, J., Ramuz, M., Liautard, J.P., The analysis of the intramacrophagic virulome of Brucella suis deciphers the environment encountered by the pathogen inside the macrophage host cell (2002) Proc. Natl. Acad. Sci. U. S. A., 99, pp. 15711-15716
  • Treptow, N.A., Shuman, H.A., Genetic evidence for substrate and periplasmic-binding-protein recognition by the MalF and MalG proteins, cytoplasmic membrane components of the Escherichia coli maltose transport system (1985) J. Bacteriol., 163, pp. 654-660
  • Wright, L., Blagova, E., Levdikov, V.M., Brannigan, J.A., Pattenden, R.J., Chambers, J., Wilkinson, A.J., Crystallization of the oligopeptidebinding protein AppA from Bacillus subtilis. Acta Crystallogr (2004) D Biol. Crystallogr., 60, pp. 175-177
  • Smith, L.T., Pocard, J.A., Bernard, T., Le Rudulier, D., Osmotic control of glycine betaine biosynthesis and degradation in Rhizobium meliloti (1988) J. Bacteriol., 170, pp. 3142-3149
  • Boch, J., Kempf, B., Bremer, E., Osmoregulation in Bacillus subtilis: synthesis of the osmoprotectant glycine betaine from exogenously provided choline (1994) J. Bacteriol., 176, pp. 5364-5371
  • van der Heide, T., Poolman, B., Osmoregulated ABC-transport system of Lactococcus lactis senses water stress via changes in the physical state of the membrane (2000) Proc. Natl. Acad. Sci. U. S. A., 97, pp. 7102-7106
  • Konstantinova, S.V., Tell, G.S., Vollset, S.E., Nygard, O., Bleie, O., Ueland, P.M., Divergent associations of plasma choline and betaine with components of metabolic syndrome in middle age and elderly men and women (2008) J. Nutr., 138, pp. 914-920
  • Lockman, P.R., Allen, D.D., The transport of choline (2002) Drug Dev. Ind. Pharm., 28, pp. 749-771
  • Pomfret, E.A., daCosta, K.A., Schurman, L.L., Zeisel, S.H., Measurement of choline and choline metabolite concentrations using high-pressure liquid chromatography and gas chromatography-mass spectrometry (1989) Anal. Biochem., 180, pp. 85-90

Citas:

---------- APA ----------
Herrmann, C.K., Bukata, L., Melli, L., Marchesini, M.I., Caramelo, J.J. & Comerci, D.J. (2013) . Identification and characterization of a high-affinity choline uptake system of Brucella abortus. Journal of Bacteriology, 195(3), 493-501.
http://dx.doi.org/10.1128/JB.01929-12
---------- CHICAGO ----------
Herrmann, C.K., Bukata, L., Melli, L., Marchesini, M.I., Caramelo, J.J., Comerci, D.J. "Identification and characterization of a high-affinity choline uptake system of Brucella abortus" . Journal of Bacteriology 195, no. 3 (2013) : 493-501.
http://dx.doi.org/10.1128/JB.01929-12
---------- MLA ----------
Herrmann, C.K., Bukata, L., Melli, L., Marchesini, M.I., Caramelo, J.J., Comerci, D.J. "Identification and characterization of a high-affinity choline uptake system of Brucella abortus" . Journal of Bacteriology, vol. 195, no. 3, 2013, pp. 493-501.
http://dx.doi.org/10.1128/JB.01929-12
---------- VANCOUVER ----------
Herrmann, C.K., Bukata, L., Melli, L., Marchesini, M.I., Caramelo, J.J., Comerci, D.J. Identification and characterization of a high-affinity choline uptake system of Brucella abortus. J. Bacteriol. 2013;195(3):493-501.
http://dx.doi.org/10.1128/JB.01929-12