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Effect of loss of clpP2, clpX and clpC1 genes on proteome of Mycobacterium tuberculosis.

ABSTRACT: The current work involves the quantitative analysis of total proteome of M. tuberculosis H37Rv depleted with the expression of different clp genes viz., clpP2, clpX and clpC1, respectively, by iTRAQ. Silencing of these genes was performed by CRISPRi approach by co-expression of specific guide sequence and dCas9 under the regulation of ATc-inducible promoter.

ORGANISM(S): Mycobacterium

SUBMITTER: Dr Nisheeth Agrawal

PROVIDER: PXD013156 | JPOST Repository | Fri Mar 20 00:00:00 GMT 2020

REPOSITORIES: jPOST

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Project description:The alternative sigma factor RpoS plays a central role in the critical host-adaptive response of the Lyme disease spirochete, Borrelia burgdorferi. We previously identified bbd18 as a negative regulator of RpoS but could not inactivate bbd18 in wild-type spirochetes. In the current study we employed an inducible bbd18 gene to demonstrate the essential nature of BBD18 for viability of wild-type spirochete viability in vitro and at a unique point in vivo. Transcriptomic analyses of BBD18 depleted cells demonstrated global induction of RpoS-dependent genes prior to lysis, with the absolute requirement for BBD18, both in vitro and in vivo, circumvented by deletion of rpoS. The increased expression of plasmid prophage genes and the presence of phage particles in the supernatants of lysing cultures indicate that RpoS regulates phage lysis-lysogeny decisions. Through this work we identify a mechanistic link between endogenous transducing prophages and the RpoS-dependent adaptive response of the Lyme disease spirochete. The alternative sigma factor RpoS plays a central role in the critical host-adaptive response of the Lyme disease spirochete, Borrelia burgdorferi. We previously identified bbd18 as a negative regulator of RpoS but could not inactivate bbd18 in wild-type spirochetes. In the current study we employed an inducible bbd18 gene to demonstrate the essential nature of BBD18 for viability of wild-type spirochete viability in vitro and at a unique point in vivo. Transcriptomic analyses of BBD18 depleted cells demonstrated global induction of RpoS-dependent genes prior to lysis, with the absolute requirement for BBD18, both in vitro and in vivo, circumvented by deletion of rpoS. The increased expression of plasmid prophage genes and the presence of phage particles in the supernatants of lysing cultures indicate that RpoS regulates phage lysis-lysogeny decisions. Through this work we identify a mechanistic link between endogenous transducing prophages and the RpoS-dependent adaptive response of the Lyme disease spirochete.

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Effect of loss of clpP2, clpX and clpC1 genes on proteome of Mycobacterium tuberculosis.

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