Unknown,Transcriptomics,Genomics,Proteomics

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Mycobacterium bovis transcriptional response to nitric oxide

ABSTRACT: We used microarray hybridization to analyze the expression pattern of M. bovis after exposure to nitric oxide. In M. tuberculosis, this treatment has been shown to induce a response similar to the hypoxic stress. Results showed the up-regulation of 51 genes belonging to the DosR regulon. The most remarkable difference with M. tuberculosis was the lack of expression of narX and narK2 in M. bovis, both genes related to survival during hypoxic stress. We further confirm this by qRT-PCR and also explore differential activity at promoter level. Four independent biological replicates were analyzed for each condition, and two dye-swap experiments were included.

ORGANISM(S): Mycobacterium bovis

SUBMITTER: German Rehren

PROVIDER: E-GEOD-15164 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Despite being one of the first eukaryotic transcriptional regulatory elements identified, the sequence of a native TATA box and its significance remain elusive. Applying criteria associated with TATA boxes we queried several Saccharomyces genomes and arrived at the consensus TATA(A/T)A(A/T)(A/G). Approximately 20% of yeast genes contain a TATA box. Strikingly, TATA box-containing genes are associated with responses to stress, are highly regulated, and preferentially utilize SAGA rather than TFIID when compared to TATA-less promoters. Transcriptional regulation in yeast appears to be mechanistically bipolar, possibly reflecting a need to balance inducible stress-related responses with constitutive housekeeping functions. A strain containing amino terminal HA-tagged TBP and its parental untagged counterpart BY4741 (Resgen) were grown at 23?C in CSM to OD600 = 0.8-1.0. Cultures were shifted to 37?C for 5 minutes (to mimic heat treatments used elsewhere (Huisinga and Pugh, 2004)), then crosslinked at 23?C with 1% formaldehyde. ChIP was performed as described previously (Strahl-Bolsinger et al., 1997) with the some modification. Following cell disruption with glass beads, the chromatin was partially purified by centrifugation (Kurdistani and Grunstein, 2003). HA-TBP was immunopurified with 12CA5 antibody. After elution and reversal of the crosslinks, samples were subjected to non-specific amplification, labeling, and array hybridization as described (Bohlander et al., 1992; Chitikila et al., 2002). The amplification procedure was modified as follows. Following 15 round B amplification cycles, DNA was purified via QIAquick PCR Purification kit. Ten cycles were used in round C. Intergenic microarrays were generated by PCR amplification of entire intergenic regions of all yeast genes, then spotted onto glass slides. Spot intensities were filtered to remove any signal that was less then one standard deviation above local background. Three replicates were performed and their log2 ratios (enriched/input) were averaged. Keywords = Chromatin Immunoprecipitation Keywords = genome-wide binding

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Mycobacterium bovis transcriptional response to nitric oxide

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