Abstract

Thioredoxins (Trxs) are antioxidant proteins that are conserved among all species. These proteins have been extensively studied and perform reducing reactions on a broad range of substrates. Here, we identified Caulobacter crescentus Trx1 (CCNA_03653; CcTrx1) as an oxidoreductase that is involved in the cell cycle progression of this model bacterium and is required to sustain life. Intriguingly, the abundance of CcTrx1 varies throughout the C. crescentus cell cycle: although the expression of CcTrx1 is induced in stalked cells, right before DNA replication initiation, CcTrx1 is actively degraded by the ClpXP protease in predivisional cells. Importantly, we demonstrated that regulation of the abundance of CcTrx1 is crucial for cell growth and survival as modulating CcTrx1 levels leads to cell death. Finally, we also report a comprehensive biochemical and structural characterization of this unique and essential Trx. The requirement to precisely control the abundance of CcTrx1 for cell survival underlines the importance of redox control for optimal cell cycle progression in C. crescentus.

Highlights

  • Thioredoxins (Trxs) are antioxidant proteins that are conserved among all species

  • The abundance of CcTrx1 varies throughout the C. crescentus cell cycle: the expression of CcTrx1 is induced in stalked cells, right before DNA replication initiation, CcTrx1 is actively degraded by the ClpXP protease in predivisional cells

  • We report the characterization of CcTrx1, a bacterial oxidoreductase that is essential for the growth and survival of C. crescentus

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Summary

ARTICLE cro

Goemans‡§¶1, François Beaufay§2, Khadija Wahni¶ʈ**, Inge Van Molle¶ʈ**3, Joris Messens¶ʈ**4, and Jean-François Collet‡§¶5 From ‡WELBIO, Avenue Hippocrate 75, 1200 Brussels, Belgium, the §de Duve Institute, Université catholique de Louvain, Avenue Hippocrate 75, 1200 Brussels, Belgium, the ¶Brussels Center for Redox Biology, 1200 Brussels, Belgium, the ʈCenter for Structural Biology, Vlaams Instituut voor Biotechnologie (VIB), 1050 Brussels, Belgium, and **Structural Biology Brussels, Vrije Universiteit Brussel, 1050 Brussels, Belgium

Edited by Ursula Jakob
Results
Protein Data Bank code
Discussion
Experimental procedures
Protein purification and antibody production
Insulin reduction assay
Redox potential
Km determination
Depletion of ClpX and ClpP
Growth curves
Full Text
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