MassIVE MSV000084267

Complete Public PXD015262

ClpX, activated by SSB, is required for DNA replication in Mycobacterium tuberculosis

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Description

The ClpP1P2 proteolytic complex is essential in Mycobacterium tuberculosis (Mtb). ClpP1P2 mediated proteolysis by ClpP1P2 requires an associated ATPase, either ClpX or ClpC1. Here, we seek to define the unique contributions of the ClpX ATPase to mycobacterial growth. We formally demonstrate that ClpX is essential for mycobacterial growth and to understand its essential functions, we identify ClpX-His-interacting proteins by pulldown and tandem mass spectrometry. We find an unexpected association between ClpX and proteins involved in DNA replication, and confirm a physical association between ClpX and the essential DNA maintenance protein Single-Stranded DNA Binding protein (SSB). Although pure SSB is not degraded by ClpXP1P2, it instead enhances ATPase hydrolysis by ClpX and degradation of the model substrate GFP-SsrA by ClpXP1P2. This activation of ClpX is mediated by the C-terminal tail of SSB that had previously been implicated in the activation of other ATPases associated with DNA replication. Consistent with the predicted interactions, depletion of ClpX perturbs DNA replication. These data reveal that ClpX serves a necessary function in DNA replication and identify the first activator of ClpX in mycobacteria. [doi:10.25345/C5ZT0G] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: ClpX ; ATPase ; ATPgammaS ; substrate

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Principal Investigators:
(in alphabetical order) 		Jemila Kester, HSPH, United States
Sarah Fortune, HSPH, USA
Submitting User: jckester
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