title: A lysozyme-like protein in Brucella abortus is involved in the early stages of intracellular replication.
abstract: Secretion of proteins in Gram-negative bacteria is a high-energy-consuming process that requires translocation across two membranes and a periplasmic space composed of a mesh-like layer, the peptidoglycan. To achieve this, bacteria have evolved complex secretion systems that cross these barriers, and in many cases there are specific peptidoglycanases that degrade the peptidoglycan to allow the proper assembly of the secretion machinery. We describe here the identification and characterization of a muramidase in Brucella abortus that participates in the intracellular multiplication in professional and nonprofessional phagocytes. We demonstrated that this protein has peptidoglycanase activity, that a strain with a clean deletion of the gene displayed a defect in the early stages of the intracellular multiplication curve, and that this is dependent on the lytic activity. While neither the attachment nor the invasion of the strain was affected, we demonstrated that it had a defect in excluding the lysosomal marker LAMP-1 but not in acquiring the reticulum endoplasmic marker calnexin, indicating that the gene participates in the early stages of the intracellular trafficking but not in the establishment of the replicative niche. Analysis of the assembly status and functionality of the VirB secretion apparatus indicated that the mutant has affected the proper function of this central virulence factor.
authors: Instituto de Investigaciones Biotecnológicas Dr Rodolfo A Ugalde, Instituto Tecnológico de Chascomús, IIB-INTECH, CONICET, Universidad Nacional de San Martín, San Martín, Buenos Aires, Argentina. jugalde@iibintech.com.ar
authors: Mariela G
authors: MG
authors: Del Giudice
authors: Juan E
authors: JE
authors: Ugalde
authors: Cecilia
authors: C
authors: Czibener
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002003:Brucella abortus / Q000166:cytology / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant; D004847:Epithelial Cells / Q000382:microbiology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D009113:Muramidase / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D010957:Plasmids
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D009113:Muramidase