Protein inactivation in mycobacteria by controlled proteolysis and its application to deplete the beta subunit of RNA polymerase.

ABSTRACT: Using a component of the Escherichia coli protein degradation machinery, we have established a system to regulate protein stability in mycobacteria. A protein tag derived from the E. coli SsrA degradation signal did not affect several reporter proteins in wild-type Mycobacterium smegmatis or Mycobacterium tuberculosis. Expression of the adaptor protein SspB, which recognizes this modified tag and helps deliver tagged proteins to the protease ClpXP, strongly decreased the activities and protein levels of different reporters. This inactivation did not occur when the function of ClpX was inhibited. Using this system, we constructed a conditional M. smegmatis knockdown mutant in which addition of anhydrotetracycline (atc) caused depletion of the beta subunit of RNA polymerase, RpoB. The impact

SUBMITTER: Kim JH 

PROVIDER: S-EPMC3064785 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature