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Brodin, Priscille,Poquet, Yannick,Levillain, Florence,Peguillet, Isabelle,Larrouy-Maumus, Gerald,Gilleron, Martine,Ewann, Fanny,Christophe, Thierry,Fenistein, Denis,Jang, Jichan,Jang, Mi-Seon,Park, Se Public Library of Science 2010 PLoS pathogens Vol.6 No.9
<▼1><P>The ability of the tubercle bacillus to arrest phagosome maturation is considered one major mechanism that allows its survival within host macrophages. To identify mycobacterial genes involved in this process, we developed a high throughput phenotypic cell-based assay enabling individual sub-cellular analysis of over 11,000 <I>Mycobacterium tuberculosis</I> mutants. This very stringent assay makes use of fluorescent staining for intracellular acidic compartments, and automated confocal microscopy to quantitatively determine the intracellular localization of <I>M. tuberculosis</I>. We characterised the ten mutants that traffic most frequently into acidified compartments early after phagocytosis, suggesting that they had lost their ability to arrest phagosomal maturation. Molecular analysis of these mutants revealed mainly disruptions in genes involved in cell envelope biogenesis (<I>fadD28</I>), the ESX-1 secretion system (<I>espL</I>/Rv3880), molybdopterin biosynthesis (<I>moaC1</I> and <I>moaD1</I>), as well as in genes from a novel locus, Rv1503c-Rv1506c. Most interestingly, the mutants in Rv1503c and Rv1506c were perturbed in the biosynthesis of acyltrehalose-containing glycolipids. Our results suggest that such glycolipids indeed play a critical role in the early intracellular fate of the tubercle bacillus. The unbiased approach developed here can be easily adapted for functional genomics study of intracellular pathogens, together with focused discovery of new anti-microbials.</P></▼1><▼2><P><B>Author Summary</B></P><P>One of the major virulence mechanisms of the tuberculosis bacillus, <I>Mycobacterium tuberculosis</I>, is its ability to resist killing by phagocytic cells of the host immune system, namely the macrophages. Macrophages degrade invading microbes by engulfment inside a vacuole, or phagosome, that progressively acidifies and accumulates hydrolytic properties. <I>M. tuberculosis</I> has the unique ability to block phagosome maturation and acidification. To identify mycobacterial genes involved in phagosome maturation arrest, we developed a novel high-throughput technology based on automated confocal microscopy. We screened a library containing over 11,000 <I>M. tuberculosis</I> mutants, and we could identify 10 mutants that had lost their ability to resist phagosome acidification. Genetic characterization of these mutants revealed that they carried lesions in genes involved in various cell processes, including biogenesis of the cell envelope. In particular, two independent mutants in the same genetic locus showed altered production of two lipids, namely diacyltrehalose (DAT) and sulfoglycolipid (SGL). <I>In vitro</I> experiments showed that SGL can indeed influence phagosome maturation. Our study unravels the role of novel lipid molecules in mycobacterial intracellular parasitism; our approach may be useful to identify virulence genes in other intracellular pathogens, and to identify novel antimicrobials.</P></▼2>
Application of Hybrid Polymeric Complexes to Solid State and Materials Chemistry
Josik Portier,Guy Campet,Nadine Treuil,Armel Poquet,Kim, Young Il,Kwon, Soon Jae,Kwak, Seo Young,Choy, Jin Ho Korean Chemical Society 1998 대한화학회지 Vol.42 No.4
A bird's-eye view on preparation, structure and properties of polymeric complexes in the field of Inorganic-Organic-Hybrids is presented in the view point of solid state and materials chemistry. These materials are useful precursors for preparing nanoparticles and fine grain oxides. Some of them are electroactive and are used as protonic or lithium electrolytes, electrochromic materials or membranes for sensors and actuators. New results on bio-hybrids, a class of material not far from polymeric complexes, are also described.