Mycobacteriophage D29 induced association of Mycobacterial RNA polymerase with ancillary factors leads to increased transcriptional activity

Abstract

Mycobacteriophage D29 infects species belonging to the genus Mycobacterium including the deadly pathogen Mycobacterium tuberculosis. D29 is a lytic phage, although, related to the lysogenic mycobacteriophage L5. This phage is unable to lysogenize in mycobacteria as it lacks the gene encoding the phage repressor. Infection by many mycobacteriophages cause various changes in the host that ultimately leads to inactivation of the latter. One of the host targets often modified in the process is RNA polymerase. During our investigations with phage D29 infected Mycobacterium smegmatis (Msm) we observed that the promoters from both phage, and to a lesser extent those of the host were found to be more active in cells that were exposed to D29, as compared to the unexposed. Further experiments indicate that the RNA polymerase purified from phage infected cells possessed higher affinity for promoters particularly those that were phage derived. Comparison of the purified RNA polymerase preparations from infected and uninfected cells showed that several ancillary transcription factors, Sigma factor F, Sigma factor H, CarD and RbpA are prominently associated with the RNA polymerase from infected cells. Based on our observations we conclude that the higher activity of RNA polymerase observed in D29 infected cells is due to its increased association with ancillary transcription factors.

Evaluation of distinct molecular architectures and coordinated regulation of the catabolic pathways of oestrogenic dioctyl phthalate isomers in Gordonia sp.

“…As described above, the harvested cells were lysed to obtain cell-free extract (CFE) preparations. The enzyme preparations were lyophilized using an Edwards lyophilizer and processed further for trypsin digestion, followed by LC-ESI-MS/MS analysis [ 32 ] using a Waters Xero-G2-Xs-QTOF system in positive ion mode. A C18 BEH column (Agilent) was used for separation chemistry using water (A) and acetonitrile (B) supplemented with 0.1 % formic acid as a mobile phase in a gradient mode.…”

Biochemical and Multi-Omics Approaches To Obtain Molecular Insights into the Catabolism of the Plasticizer Benzyl Butyl Phthalate in Rhodococcus sp. Strain PAE-6

“…For proteome analysis, cell extracts as mentioned above, were lyophilized using an Edwards lyophilizer and processed further for trypsin digestion, followed by ESI-LC-MS ( 61 ). A C 18 BEH column (Agilent) was used for peptide separation, and a Waters Xero-G2-Xs-QTof mass spectrometer was used in positive ion mode for analyzing the peptides.…”

The respiratory lipoquinone, menaquinone, functions as an inducer of genes regulated by the Mycobacterium smegmatis repressor MSMEG_2295

“…A 144 bp DNA fragment [24] harbouring the MSMEG_2295 binding site upstream of the dinB2 operon was amplified using the primers P tetR fw Cy5 and P tetR rv (Table S3). The Cy5-labelled fragment was subjected to a binding reaction using different concentrations of Ni-NTA agarose purified hexa-His-tagged MSMEG_2295 protein isolated as described in our earlier study [12].…”