Subcellular Localization of the Intracellular Survival-Enhancing Eis Protein of
            <i>Mycobacterium tuberculosis</i>

Dahl, John L.; Wei, Jun; Moulder, James W.; Laal, Suman; Friedman, Richard L.

doi:10.1128/iai.69.7.4295-4302.2001

Cited by 43 publications

(46 citation statements)

References 60 publications

(51 reference statements)

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“…Coverslips were again washed, and incubated with a 1 : 500 dilution of anti-Eis antibody for 1 h at room temperature on an orbital shaker. Anti-Eis antibody is an affinity-purified antibody to Eis generated in rabbits as described previously (Dahl et al, 2001). Alexafluor 594 goat antirabbit IgG linked to a fluorescent red dye (Molecular Probes;…”

Section: Methodsmentioning

confidence: 99%

“…Because the Eis protein was shown to be secreted into the bacterial culture medium by M. tuberculosis in vitro (Dahl et al, 2001), studies were done to examine infected macrophages for the presence of the Eis protein in the host cell cytoplasm. M. tuberculosis H37Ra and Deis strains were transformed with the pBEN plasmid, which contains the green fluorescent protein (gfp) gene driven by phsp60 from Mycobacterium bovis BCG (Saviola et al, 2003).…”

Section: Presence Of Eis In the Cytoplasm Of Macrophages Infected Witmentioning

confidence: 99%

“…M. tuberculosis H37Ra wild-type and Deis strains expressing gfp were named MtbRa/gfp and MtbRaDeis/gfp, respectively. The MtbRa/gfp-and MtbRaDeis/gfp-infected cells were washed, permeabilized, and probed with rabbit anti-Eis antibody (Dahl et al, 2001) followed by goat anti-rabbit antibody labelled with a fluorescent red dye.…”

Section: Presence Of Eis In the Cytoplasm Of Macrophages Infected Witmentioning

confidence: 99%

“…As the anti-Eis antibody used in these studies is affinity purified (Dahl et al, 2001), it is unlikely to cross-react with host cell proteins. Eis protein has a tendency to aggregate in solution and this band is probably the result of aggregate formation.…”

Section: Western Blot Analysis Of the Cytoplasm Of Infected Macrophagmentioning

confidence: 99%

“…The Eis protein is a 42 kDa protein that is secreted when M. tuberculosis is grown in liquid culture (Dahl et al, 2001). In the present study, we constructed and utilized an eis mutant (Deis) and complemented strains of M. tuberculosis H37Ra and H37Rv to study the role the gene plays in the microbe's interactions with macrophages.…”

Section: Introductionmentioning

confidence: 99%

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Expression, production and release of the Eis protein by Mycobacterium tuberculosis during infection of macrophages and its effect on cytokine secretion

et al. 2007

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The eis gene of Mycobacterium tuberculosis has been shown to play a role in the survival of the avirulent Mycobacterium smegmatis within the macrophage. In vitro and in vivo analysis of Deis deletion mutants and complemented strains showed no effect on survival of M. tuberculosis in U-937 macrophages or in a mouse aerosol infection model, respectively. Further studies were done in an attempt to determine the role of eis in M. tuberculosis intracellular survival and to define a phenotypic difference between wild-type and the Deis deletion mutant. Bioinformatic analysis indicated that Eis is an acetyltransferase of the GCN5-related family of N-acetyltransferases. Immunofluorescence microscopy and Western blot analysis studies demonstrated that Eis is released into the cytoplasm of M. tuberculosis-infected U-937 macrophages. Eis was also found in the extravesicular fraction and culture supernatant of M. tuberculosis-infected macrophages. The effect of Eis on human macrophage cytokine secretion was also examined. Eis modulated the secretion of IL-10 and TNF-a by primary human monocytes in response both to infection with M. tuberculosis and to stimulation with recombinant Eis protein. These results suggest that Eis is a mycobacterial effector that is released into the host cell to modulate inflammatory responses, possibly via transcriptional or post-translational means.

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Section: Methodsmentioning

confidence: 99%

Section: Presence Of Eis In the Cytoplasm Of Macrophages Infected Witmentioning

confidence: 99%

Section: Presence Of Eis In the Cytoplasm Of Macrophages Infected Witmentioning

confidence: 99%

Section: Western Blot Analysis Of the Cytoplasm Of Infected Macrophagmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression, production and release of the Eis protein by Mycobacterium tuberculosis during infection of macrophages and its effect on cytokine secretion

et al. 2007

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Identification and Characterization of Inhibitors of the Aminoglycoside Resistance Acetyltransferase Eis from Mycobacterium tuberculosis

2011

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Association of the Rv0679c protein with lipids and carbohydrates in Mycobacterium tuberculosis/Mycobacterium bovis BCG

2007

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The Rv0679c gene in Mycobacterium tuberculosis H37Rv encodes a protein with a predicted molecular mass of 16,586 Da consisting of 165 amino acids which contains a putative N-terminal signal sequence and a consensus lipoprotein-processing motif. Globomycin treatment, Triton X-114 separation and mass spectrometry analyses clarified a property of the Rv0679c protein as a lipoprotein. In addition, trifluoromethanesulphonic acid treatment of the lysate revealed an association of the recombinant Rv0679c protein with carbohydrates. The Rv0679c protein homolog of Mycobacterium bovis BCG was also expressed as the protein associated with lipids and carbohydrates. In Western blot analysis, each of the protein homolog and Lipoarabinomannan (LAM) was detected as a similar pattern by anti-Rv0679c and anti-LAM antibodies, respectively. Interestingly, the Rv0679c protein was detected in commercially available LAM purified from M. tuberculosis. Inhibition assay of LAM synthesis in M. bovis BCG by ethambutol showed an altered migration pattern of the Rv0679c protein to low molecular mass similar to that of LAM. The results suggest that the Rv0679c protein exists as a tight complex with LAM in M. tuberculosis/M. bovis BCG.

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Subcellular Localization of the Intracellular Survival-Enhancing Eis Protein of Mycobacterium tuberculosis

Cited by 43 publications

References 60 publications

Expression, production and release of the Eis protein by Mycobacterium tuberculosis during infection of macrophages and its effect on cytokine secretion

Expression, production and release of the Eis protein by Mycobacterium tuberculosis during infection of macrophages and its effect on cytokine secretion

Identification and Characterization of Inhibitors of the Aminoglycoside Resistance Acetyltransferase Eis from Mycobacterium tuberculosis

Association of the Rv0679c protein with lipids and carbohydrates in Mycobacterium tuberculosis/Mycobacterium bovis BCG

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