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The Mycobacterium tuberculosis Rv2745c Plays an Important Role in Responding to Redox Stress

ABSTRACT: The Rv2745c (clgR) gene encodes a Clp protease gene regulator that is induced in response to a variety of stress conditions and potentially plays a role in Mtb pathogenesis. The isogenic Mtb:ΔRv2745c mutant is significantly more sensitive to in vitro redox stress generated by diamide, relative to wild-type Mtb, implicating a role for ClgR in the management of intraphagosomal redox stress. Redox stress led to dysregulation of the σH regulon in the isogenic mutant, Mtb:ΔRv2745c. Induction of clgR in Mtb and Mtb:ΔRv2745c (comp) did not lead to Clp protease induction, indicating that clgR has additional functions that need to be elucidated. Disruption of genes involved in sulfate assimilation also occurred in the knock out, implicating clgR as a possible regulator of downstream signaling cascades that facilitate Mtb survival.

ORGANISM(S): Mycobacterium tuberculosis H37Rv Mycobacterium tuberculosis CDC1551

PROVIDER: GSE55817 | GEO | 2014/03/25

SECONDARY ACCESSION(S): PRJNA242600

REPOSITORIES: GEO

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The Mycobacterium tuberculosis Rv2745c Plays an Important Role in Responding to Redox Stress

Project description:The Rv2745c (clgR) gene encodes a Clp protease gene regulator that is induced in response to a variety of stress conditions and potentially plays a role in Mtb pathogenesis. The isogenic Mtb:ΔRv2745c mutant is significantly more sensitive to in vitro redox stress generated by diamide, relative to wild-type Mtb, implicating a role for ClgR in the management of intraphagosomal redox stress. Redox stress led to dysregulation of the σH regulon in the isogenic mutant, Mtb:ΔRv2745c. Induction of clgR in Mtb and Mtb:ΔRv2745c (comp) did not lead to Clp protease induction, indicating that clgR has additional functions that need to be elucidated. Disruption of genes involved in sulfate assimilation also occurred in the knock out, implicating clgR as a possible regulator of downstream signaling cascades that facilitate Mtb survival. At different time points, (30, 60 and 90 mins, t=30, t=60 and t=90) following the addition of diamide to M. tuberculosis CDC1551 cultures, RNA was extracted and labeled with Cy5. RNA was also extracted from pre-diamide addition (t=0) time points and labeled with Cy3. These samples were cohybridized on M. tuberculosis CDC1551 whole genome microarrays using biological triplicate samples (i.e. samples derived from three distinct cultures and treatments) and thus three unique datasets were generated for each of the three time points for Mtb. Similar experiments were performed for an isogenic Rv2745c (clgR) mutant in Mtb CDC1551 which was generated by allelic exchange and a complemented strain which was generated by trans-complementation of Rv2745c in the attB locus of the mutant strain, thus restoring the wild type regulation of Rv2745c.

2014-03-25 | E-GEOD-55817 | biostudies-arrayexpress

The Mycobacterium tuberculosis Clp Gene Regulator is Required for in vitro Reactivation from Hypoxia-induced Dormancy

2014-12-12 | GSE64065 | GEO

The Mycobacterium tuberculosis Clp Gene Regulator is Required for in vitro Reactivation from Hypoxia-induced Dormancy

Project description:The isogenic Mtb:ΔRv2745c mutant is significantly more sensitive normoxia conditions after a period of hypoxia, relative to wild-type Mtb, implicating a role for ClgR in response to reactivation, in vivo. Both hypoxia and reaeration treatment led to dysregulation of the σH regulon in the isogenic mutant, Mtb:ΔRv2745c. Induction of clgR in Mtb did lead to Clp protease induction, indicating that clgR plays a role in differntially activating downstream genes in a condition dependent manner. Disruption of genes involved in the dosR regulon, the Enduring Hypoxia Response, lipid synthesis, and mycolic acid synthesis also occurred in the knock out, implicating clgR as a possible regulator of downstream signaling cascades that facilitate Mtb survival. There was also a differnetial response of genes that are known Clp protease targets, in addition to potential Clp protease targets that had similar induction patterns as known Clp protease targets. At different time points, following hypoxia (day 1, day 5, and day 7) and reaeration (1 hour, 4 hour, 6 hour, 12 hour, 24 hour, and 48 hour) treatments M. tuberculosis CDC1551 cultures, RNA was extracted and labeled with Cy5. RNA was also extracted from pre-hypoxia (t=0) time points and labeled with Cy3. These samples were cohybridized on M. tuberculosis CDC1551 whole genome microarrays using biological triplicate samples (i.e. samples derived from three distinct cultures and treatments) and thus three unique datasets were generated for each of the three time points for Mtb. Similar experiments were performed for an isogenic Rv2745c (clgR) mutant in Mtb CDC1551 which was generated by allelic exchange.

2014-12-12 | E-GEOD-64065 | biostudies-arrayexpress

Functional Genomics Reveals Extended Roles of the Mycobacterium tuberculosis Stress Response Factor sigmaH

2009-09-04 | GSE15608 | GEO

Functional Genomics Reveals Extended Roles of the Mycobacterium tuberculosis Stress Response Factor sigmaH

Project description:DNA Microarray based comparison of M. tuberculosis and its isogenic delta-sigmaH mutant for an extended period of time, using a diamide-stress and removal strategy (post-diamide stress or PDS) Raw data has been requested of, but not provided by, the submitter. Two strains, M. tuberculosis vs delta-sigmaH mutant. At Pre-diamide (control), 0' 30' 60' 90' and 120' post-diamide stress, two biological replicates, six replicates per array

2010-06-24 | E-GEOD-15608 | biostudies-arrayexpress

Sigma factor B (sigB) response to stress conditions (0.05% SDS and 5mM Diamide)

Project description:The numerous sigma factors present in Mycobacterium tuberculosis (MTB) are indicative of adaptability to different environmental conditions. In this report we describe the sigma factor B (sigB) regulon and the phenotypes of a MTB sigB mutant strain exposed to different stresses like SDS and Diamide. This experiment set compares expression profiles between H37Rv wild type and H37Rv sigB null mutant as well as under different stress conditions. Both H37Rv wild type and H37Rv sigB null mutants were treated with either 0.05% SDS or 5mM Diamide for 60 min and their expression profiles were compared with untreated wild type or mutant controls. Biological Replicate

2010-06-25 | E-GEOD-15976 | biostudies-arrayexpress

Sigma factor B (sigB) response to stress conditions (0.05% SDS and 5mM Diamide)

2009-06-23 | GSE15976 | GEO

Global proteomic analysis for stress response of Mycobacterium tuberculosis

Project description:To expand an insight into the stress adaptation in Mycobacterium tuberculosis H37Rv (Mtb), we approached the iTRAQ analysis to understand the global proteome profile of stressed Mtb under Acid (5.5 pH), Diamide (5mM) and Hydrogen peroxide (H2O2) (5mM) conditions and compared with gene expression of Mtb incubated in 7H9 rich medium (pH 7.0). The experiment was performed for overnight treatment of aforementioned stress. Whole cell lysate (WCL) from three independent repeats of control and stressed samples were labelled with three set of 4-plex iTRAQ labelling reagents (AB SCIEX) in proportion to manufacturer’s protocol.

2022-10-13 | PXD015167 | Pride

Reaeration timecourse from a defined hypoxia model

Project description:Reaeration timecourse from a defined hypoxia model in Mycobacterium tuberculosis. The mechanism by which mycobacteria return to a replicating state upon re-exposure to favorable conditions is not well understood. In this study, we utilized reaeration from a defined hypoxia model to characterize the adaptive response of Mycobacterium tuberculosis (MTB) following a return to favorable growth conditions. Global transcriptional analysis identified the ~100 gene Reaeration Response, induced relative to both log-phase and hypoxic MTB. This response includes chaperones and proteases, as well as the transcription factor Rv2745c, a Clp protease gene regulator (ClgR) orthologue. During reaeration, genes repressed during hypoxia are also upregulated in a wave of transcription that includes genes crucial to transcription, translation and oxidative phosphorylation. Each reaerating culture compared at multiple timepoints to a sample from the same culture at seven days of hypoxia. Three or more replicates at each timepoint.

2010-07-28 | E-GEOD-21590 | biostudies-arrayexpress

Reaeration timecourse from a defined hypoxia model

2010-07-28 | GSE21590 | GEO

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