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Identification of a proteolysis regulator for an essential enzyme in Mycobacterium tuberculosis

Kahne, S. C.; Yoo, J. H.; Chen, J.; Nakedi, K.; Iyer, L. M.; Putzel, G.; Samhadaneh, N. M.; Pironti, A.; Aravind, L.; Ekiert, D. C.; Bhabha, G.; Rhee, K. Y.; Darwin, K. H.

2024-03-30 microbiology
10.1101/2024.03.29.587195 bioRxiv
Show abstract

In Mycobacterium tuberculosis proteins that are post-translationally modified with Pup, a prokaryotic ubiquitin-like protein, can be degraded by proteasomes. While pupylation is reversible, mechanisms regulating substrate specificity have not been identified. Here, we identify the first depupylation regulators: CoaX, a pseudokinase, and pantothenate, an essential, central metabolite. In a {Delta}coaX mutant, pantothenate synthesis enzymes were more abundant, including PanB, a substrate of the Pup-proteasome system. Media supplementation with pantothenate decreased PanB levels in a coaX and Pup-proteasome-dependent manner. In vitro, CoaX accelerated depupylation of Pup[~]PanB, while addition of pantothenate inhibited this reaction. Collectively, we propose CoaX contributes to proteasomal degradation of PanB by modulating depupylation of Pup[~]PanB in response to pantothenate levels. One Sentence SummaryA pseudo-pantothenate kinase regulates proteasomal degradation of a pantothenate synthesis enzyme in M. tuberculosis.

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