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Identification of Lpta, Lpxe, and Lpxo, Three Genes

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Identification of Lpta, Lpxe, and Lpxo, Three Genes Identification of lptA, lpxE, and lpxO, Three Genes Involved in the Remodeling of Brucella Cell Envelope Raquel Conde-Alvarez, Leyre Palacios-Chaves, Yolanda Gil-Ramirez, Miriam Salvador-Bescos, Marina Barcena-Varela, Beatriz Aragon-Aranda, Estrella Martinez-Gomez, Amaia Zuniga-Ripa, Maria J. Miguel, Toby Leigh Bartholomew, et al. To cite this version: Raquel Conde-Alvarez, Leyre Palacios-Chaves, Yolanda Gil-Ramirez, Miriam Salvador-Bescos, Ma- rina Barcena-Varela, et al.. Identification of lptA, lpxE, and lpxO, Three Genes Involved inthe Remodeling of Brucella Cell Envelope. Frontiers in Microbiology, Frontiers Media, 2018, 8, pp.2657. 10.3389/fmicb.2017.02657. hal-02118051 HAL Id: hal-02118051 https://hal.archives-ouvertes.fr/hal-02118051 Submitted on 21 Nov 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License fmicb-08-02657 January 9, 2018 Time: 16:56 # 1 ORIGINAL RESEARCH published: 10 January 2018 doi: 10.3389/fmicb.2017.02657 Identification of lptA, lpxE, and lpxO, Three Genes Involved in the Remodeling of Brucella Cell Envelope Raquel Conde-Álvarez1, Leyre Palacios-Chaves2, Yolanda Gil-Ramírez1, Miriam Salvador-Bescós1, Marina Bárcena-Varela1, Beatriz Aragón-Aranda1, Estrella Martínez-Gómez1, Amaia Zúñiga-Ripa1, María J. de Miguel3, Toby Leigh Bartholomew4, Sean Hanniffy5, María-Jesús Grilló2, Miguel Ángel Vences-Guzmán6, José A. Bengoechea4, Vilma Arce-Gorvel5, Jean-Pierre Gorvel5, Ignacio Moriyón1* and Maite Iriarte1* Edited by: 1 Universidad de Navarra, Facultad de Medicina, Departamento de Microbiología y Parasitología, Instituto de Salud Tropical Axel Cloeckaert, (ISTUN) e Instituto de Investigación Sanitaria de Navarra (IdISNA), Pamplona, Spain, 2 Instituto de Agrobiotecnología, Institut National de la Recherche Consejo Superior de Investigaciones Científicas – Universidad Pública de Navarra – Gobierno de Navarra, Pamplona, Spain, Agronomique (INRA), France 3 Unidad de Producción y Sanidad Animal, Instituto Agroalimentario de Aragón, Centro de Investigación y Tecnología Reviewed by: Agroalimentaria de Aragón – Universidad de Zaragoza, Zaragoza, Spain, 4 Wellcome-Wolfson Institute for Experimental Diego J. Comerci, Medicine, Queen’s University Belfast, Belfast, United Kingdom, 5 Institut National de la Santé et de la Recherche Médicale, Instituto de Investigaciones U1104, Centre National de la Recherche Scientifique UMR7280, Centre d’Immunologie de Marseille-Luminy, Aix-Marseille Biotecnológicas (IIB-INTECH), University UM2, Marseille, France, 6 Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Argentina Cuernavaca, Mexico Roy Martin Roop II, East Carolina University, United States The brucellae are facultative intracellular bacteria that cause a worldwide extended *Correspondence: Ignacio Moriyón zoonosis. One of the pathogenicity mechanisms of these bacteria is their ability to [email protected] avoid rapid recognition by innate immunity because of a reduction of the pathogen- Maite Iriarte [email protected] associated molecular pattern (PAMP) of the lipopolysaccharide (LPS), free-lipids, and other envelope molecules. We investigated the Brucella homologs of lptA, lpxE, and Specialty section: lpxO, three genes that in some pathogens encode enzymes that mask the LPS This article was submitted to Infectious Diseases, PAMP by upsetting the core-lipid A charge/hydrophobic balance. Brucella lptA, which a section of the journal encodes a putative ethanolamine transferase, carries a frame-shift in B. abortus but Frontiers in Microbiology not in other Brucella spp. and phylogenetic neighbors like the opportunistic pathogen Received: 26 October 2017 Ochrobactrum anthropi. Consistent with the genomic evidence, a B. melitensis lptA Accepted: 20 December 2017 Published: 10 January 2018 mutant lacked lipid A-linked ethanolamine and displayed increased sensitivity to Citation: polymyxin B (a surrogate of innate immunity bactericidal peptides), while B. abortus Conde-Álvarez R, Palacios-Chaves L, carrying B. melitensis lptA displayed increased resistance. Brucella lpxE encodes a Gil-Ramírez Y, Salvador-Bescos M, Bárcena-Varela M, Aragón-Aranda B, putative phosphatase acting on lipid A or on a free-lipid that is highly conserved in all Martínez-Gómez E, Zúñiga-Ripa A, brucellae and O. anthropi. Although we found no evidence of lipid A dephosphorylation, de Miguel MJ, Bartholomew TL, a B. abortus lpxE mutant showed increased polymyxin B sensitivity, suggesting the Hanniffy S, Grilló M-J, Vences-Guzmán MÁ, existence of a hitherto unidentified free-lipid involved in bactericidal peptide resistance. Bengoechea JA, Arce-Gorvel V, Gene lpxO putatively encoding an acyl hydroxylase carries a frame-shift in all brucellae Gorvel J-P, Moriyón I and Iriarte M (2018) Identification of lptA, lpxE, except B. microti and is intact in O. anthropi. Free-lipid analysis revealed that lpxO and lpxO, Three Genes Involved corresponded to olsC, the gene coding for the ornithine lipid (OL) acyl hydroxylase in the Remodeling of Brucella Cell active in O. anthropi and B. microti, while B. abortus carrying the olsC of O. anthropi Envelope. Front. Microbiol. 8:2657. doi: 10.3389/fmicb.2017.02657 and B. microti synthesized hydroxylated OLs. Interestingly, mutants in lptA, lpxE, or olsC Frontiers in Microbiology| www.frontiersin.org 1 January 2018| Volume 8| Article 2657 fmicb-08-02657 January 9, 2018 Time: 16:56 # 2 Conde-Álvarez et al. Genes Remodeling Brucella Cell Envelope were not attenuated in dendritic cells or mice. This lack of an obvious effect on virulence together with the presence of the intact homolog genes in O. anthropi and B. microti but not in other brucellae suggests that LptA, LpxE, or OL b-hydroxylase do not significantly alter the PAMP properties of Brucella LPS and free-lipids and are therefore not positively selected during the adaptation to intracellular life. Keywords: lipopolysaccharide, Brucella, lipids, cell envelope, PAMP INTRODUCTION is a eukaryotic-type phospholipid required for Brucella full virulence (Comerci et al., 2006; Conde-Alvarez et al., 2006). Brucellosis is the collective name of a group of zoonotic Among the amino lipids, only the ornithine lipids (OL) have diseases afflicting a wide range of domestic and wild mammals been investigated which unlike their counterparts in Bordetella, (Whatmore, 2009; Zheludkov and Tsirelson, 2010). In domestic do not trigger the release of IL-6 or TNF-a by macrophages, livestock brucellosis is manifested mostly as abortions and possibly on account of their longer acyl chains that reduce the OL infertility, and contact with infected animals and consumption PAMP (Palacios-Chaves et al., 2011). Concerning the LPS, most of unpasteurized dairy products are the sources of human bacteria carry C1 and C40 glucosamine disaccharides with C12 brucellosis, an incapacitating condition that requires prolonged and C14 acyl and acyl-oxyacyl chains. This highly amphipathic antibiotic treatment (Zinsstag et al., 2011). Eradicated in a structure, named lipid A, is adjacent to additional negatively handful of countries, brucellosis is endemic or even increasing in charged groups of the core oligosaccharide, namely the heptose many areas of the world (Jones et al., 2013; Ducrotoy et al., 2017; phosphates and 2-keto-3-deoxyoctulosonate carboxyl groups Lai et al., 2017). (Kastowsky et al., 1992; Moriyón, 2003). This lipid A-core PAMP This disease is caused by facultative intracellular parasites is so efficiently detected by the innate immunity system that some of the genus Brucella. Taxonomically placed in the a-2 pathogens partially conceal it by removing phosphate groups or Proteobacteria (Moreno et al., 1990), the brucellae are close substituting them with arabinosamine and/or ethanolamine, or to plant pathogens and endosymbionts such as Agrobacterium, by hydroxylating the acyl chains (Takahashi et al., 2008; Lewis Sinorhizobium, and Rhizobium and to soil bacteria such et al., 2013; Moreira et al., 2013; Needham and Trent, 2013; Llobet as Ochrobactrum, the latter including some opportunistic et al., 2015; Trombley et al., 2015). In contrast, Brucella lipid pathogens, and comparative analyses suggest that soil bacteria A is a diaminoglucose disaccharide amide-linked to long (C16, of this group are endowed with properties that represent a first C18) and very long (C28–C30) acyl chains (Velasco et al., 2000; scaffold on which an intracellular life style develops (Velasco Iriarte et al., 2004; Fontana et al., 2016). Furthermore, negative et al., 2000; Moreno and Moriyón, 2007; Barquero-Calvo et al., charges in lipid A phosphates and 2-keto-3-deoxyoctulosonate 2009). The brucellae owe their pathogenicity mainly to their are counterbalanced by four glucosamine units present in the ability to multiply within dendritic cells, macrophages, and a core (Kubler-Kielb and Vinogradov, 2013; Fontana et al., 2016). variety of other cells. Due to their ability to control intracellular As illustrated by the unusually reduced endotoxicity of the
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