author_facet Polesky, Andrea H.
Ross, Julianna T. D.
Falkow, Stanley
Tompkins, Lucy S.
Polesky, Andrea H.
Ross, Julianna T. D.
Falkow, Stanley
Tompkins, Lucy S.
author Polesky, Andrea H.
Ross, Julianna T. D.
Falkow, Stanley
Tompkins, Lucy S.
spellingShingle Polesky, Andrea H.
Ross, Julianna T. D.
Falkow, Stanley
Tompkins, Lucy S.
Infection and Immunity
Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
Infectious Diseases
Immunology
Microbiology
Parasitology
author_sort polesky, andrea h.
spelling Polesky, Andrea H. Ross, Julianna T. D. Falkow, Stanley Tompkins, Lucy S. 0019-9567 1098-5522 American Society for Microbiology Infectious Diseases Immunology Microbiology Parasitology http://dx.doi.org/10.1128/iai.69.2.977-987.2001 <jats:title>ABSTRACT</jats:title> <jats:p> <jats:italic>Legionella pneumophila</jats:italic> is a facultative intracellular gram-negative rod that causes pneumonia in humans. Free-living amoebas are thought to serve as a reservoir for <jats:italic>Legionella</jats:italic> infections. Signature-tagged mutagenesis was employed to identify <jats:italic>Legionella pneumophila</jats:italic> genes necessary for survival in the amoeba <jats:italic>Acanthamoeba castellanii</jats:italic> . Six mutant strains were defective in assays of invasion and intracellular growth. Four mutants also exhibited invasion and replication defects in <jats:italic>Hartmannella vermiformis</jats:italic> , an amoeba linked to hospital outbreaks of <jats:italic>Legionella</jats:italic> pneumonia. The six mutants also were tested in macrophages derived from peripheral blood mononuclear cells. Two mutants had intracellular replication defects, and two different strains entered cells less efficiently. Two transposon insertions were in known <jats:italic>L. pneumophila</jats:italic> genes, <jats:italic>lspK</jats:italic> and <jats:italic>aroB</jats:italic> . The other four were in novel genes. One gene has similarity to a cytochrome <jats:italic>c</jats:italic> -type biogenesis protein of <jats:italic>Pseudomonas fluorescens</jats:italic> . Another has similarity to a transcriptional activator regulating flagellar biosynthesis in <jats:italic>Vibrio cholera</jats:italic> . The third is similar to <jats:italic>traA</jats:italic> of <jats:italic>Rhizobium</jats:italic> sp. strain NGR234, which is involved in conjugal transfer of DNA. The fourth has no homology. By using survival in amoeba as a selection, we have isolated mutant strains with a range of phenotypes; and we have potentially identified new <jats:italic>L. pneumophila</jats:italic> virulence genes. </jats:p> Identification of <i>Legionella pneumophila</i> Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis Infection and Immunity
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title Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_unstemmed Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_full Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_fullStr Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_full_unstemmed Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_short Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_sort identification of <i>legionella pneumophila</i> genes important for infection of amoebas by signature-tagged mutagenesis
topic Infectious Diseases
Immunology
Microbiology
Parasitology
url http://dx.doi.org/10.1128/iai.69.2.977-987.2001
publishDate 2001
physical 977-987
description <jats:title>ABSTRACT</jats:title> <jats:p> <jats:italic>Legionella pneumophila</jats:italic> is a facultative intracellular gram-negative rod that causes pneumonia in humans. Free-living amoebas are thought to serve as a reservoir for <jats:italic>Legionella</jats:italic> infections. Signature-tagged mutagenesis was employed to identify <jats:italic>Legionella pneumophila</jats:italic> genes necessary for survival in the amoeba <jats:italic>Acanthamoeba castellanii</jats:italic> . Six mutant strains were defective in assays of invasion and intracellular growth. Four mutants also exhibited invasion and replication defects in <jats:italic>Hartmannella vermiformis</jats:italic> , an amoeba linked to hospital outbreaks of <jats:italic>Legionella</jats:italic> pneumonia. The six mutants also were tested in macrophages derived from peripheral blood mononuclear cells. Two mutants had intracellular replication defects, and two different strains entered cells less efficiently. Two transposon insertions were in known <jats:italic>L. pneumophila</jats:italic> genes, <jats:italic>lspK</jats:italic> and <jats:italic>aroB</jats:italic> . The other four were in novel genes. One gene has similarity to a cytochrome <jats:italic>c</jats:italic> -type biogenesis protein of <jats:italic>Pseudomonas fluorescens</jats:italic> . Another has similarity to a transcriptional activator regulating flagellar biosynthesis in <jats:italic>Vibrio cholera</jats:italic> . The third is similar to <jats:italic>traA</jats:italic> of <jats:italic>Rhizobium</jats:italic> sp. strain NGR234, which is involved in conjugal transfer of DNA. The fourth has no homology. By using survival in amoeba as a selection, we have isolated mutant strains with a range of phenotypes; and we have potentially identified new <jats:italic>L. pneumophila</jats:italic> virulence genes. </jats:p>
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author Polesky, Andrea H., Ross, Julianna T. D., Falkow, Stanley, Tompkins, Lucy S.
author_facet Polesky, Andrea H., Ross, Julianna T. D., Falkow, Stanley, Tompkins, Lucy S., Polesky, Andrea H., Ross, Julianna T. D., Falkow, Stanley, Tompkins, Lucy S.
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description <jats:title>ABSTRACT</jats:title> <jats:p> <jats:italic>Legionella pneumophila</jats:italic> is a facultative intracellular gram-negative rod that causes pneumonia in humans. Free-living amoebas are thought to serve as a reservoir for <jats:italic>Legionella</jats:italic> infections. Signature-tagged mutagenesis was employed to identify <jats:italic>Legionella pneumophila</jats:italic> genes necessary for survival in the amoeba <jats:italic>Acanthamoeba castellanii</jats:italic> . Six mutant strains were defective in assays of invasion and intracellular growth. Four mutants also exhibited invasion and replication defects in <jats:italic>Hartmannella vermiformis</jats:italic> , an amoeba linked to hospital outbreaks of <jats:italic>Legionella</jats:italic> pneumonia. The six mutants also were tested in macrophages derived from peripheral blood mononuclear cells. Two mutants had intracellular replication defects, and two different strains entered cells less efficiently. Two transposon insertions were in known <jats:italic>L. pneumophila</jats:italic> genes, <jats:italic>lspK</jats:italic> and <jats:italic>aroB</jats:italic> . The other four were in novel genes. One gene has similarity to a cytochrome <jats:italic>c</jats:italic> -type biogenesis protein of <jats:italic>Pseudomonas fluorescens</jats:italic> . Another has similarity to a transcriptional activator regulating flagellar biosynthesis in <jats:italic>Vibrio cholera</jats:italic> . The third is similar to <jats:italic>traA</jats:italic> of <jats:italic>Rhizobium</jats:italic> sp. strain NGR234, which is involved in conjugal transfer of DNA. The fourth has no homology. By using survival in amoeba as a selection, we have isolated mutant strains with a range of phenotypes; and we have potentially identified new <jats:italic>L. pneumophila</jats:italic> virulence genes. </jats:p>
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spelling Polesky, Andrea H. Ross, Julianna T. D. Falkow, Stanley Tompkins, Lucy S. 0019-9567 1098-5522 American Society for Microbiology Infectious Diseases Immunology Microbiology Parasitology http://dx.doi.org/10.1128/iai.69.2.977-987.2001 <jats:title>ABSTRACT</jats:title> <jats:p> <jats:italic>Legionella pneumophila</jats:italic> is a facultative intracellular gram-negative rod that causes pneumonia in humans. Free-living amoebas are thought to serve as a reservoir for <jats:italic>Legionella</jats:italic> infections. Signature-tagged mutagenesis was employed to identify <jats:italic>Legionella pneumophila</jats:italic> genes necessary for survival in the amoeba <jats:italic>Acanthamoeba castellanii</jats:italic> . Six mutant strains were defective in assays of invasion and intracellular growth. Four mutants also exhibited invasion and replication defects in <jats:italic>Hartmannella vermiformis</jats:italic> , an amoeba linked to hospital outbreaks of <jats:italic>Legionella</jats:italic> pneumonia. The six mutants also were tested in macrophages derived from peripheral blood mononuclear cells. Two mutants had intracellular replication defects, and two different strains entered cells less efficiently. Two transposon insertions were in known <jats:italic>L. pneumophila</jats:italic> genes, <jats:italic>lspK</jats:italic> and <jats:italic>aroB</jats:italic> . The other four were in novel genes. One gene has similarity to a cytochrome <jats:italic>c</jats:italic> -type biogenesis protein of <jats:italic>Pseudomonas fluorescens</jats:italic> . Another has similarity to a transcriptional activator regulating flagellar biosynthesis in <jats:italic>Vibrio cholera</jats:italic> . The third is similar to <jats:italic>traA</jats:italic> of <jats:italic>Rhizobium</jats:italic> sp. strain NGR234, which is involved in conjugal transfer of DNA. The fourth has no homology. By using survival in amoeba as a selection, we have isolated mutant strains with a range of phenotypes; and we have potentially identified new <jats:italic>L. pneumophila</jats:italic> virulence genes. </jats:p> Identification of <i>Legionella pneumophila</i> Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis Infection and Immunity
spellingShingle Polesky, Andrea H., Ross, Julianna T. D., Falkow, Stanley, Tompkins, Lucy S., Infection and Immunity, Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis, Infectious Diseases, Immunology, Microbiology, Parasitology
title Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_full Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_fullStr Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_full_unstemmed Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_short Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
title_sort identification of <i>legionella pneumophila</i> genes important for infection of amoebas by signature-tagged mutagenesis
title_unstemmed Identification of Legionella pneumophila Genes Important for Infection of Amoebas by Signature-Tagged Mutagenesis
topic Infectious Diseases, Immunology, Microbiology, Parasitology
url http://dx.doi.org/10.1128/iai.69.2.977-987.2001