Book/Dissertation / PhD Thesis FZJ-2019-03124

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Regulation and assembly of the cytochrome bc1 -aa3 supercomplex in Corynebacterium glutamicum



2019
Forschungszentrum Jülich GmbH Zentralbibliothek Verlag Jülich
ISBN: 978-3-95806-416-4

Jülich : Forschungszentrum Jülich GmbH Zentralbibliothek Verlag, Schriften des Forschungszentrums Jülich. Reihe Schlüsseltechnologien / Key Technologies 199, 135 S. () = Dissertation, Heinrich-Heine-Universität Düsseldorf, 2019

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Abstract: The synthesis of aerobic respiratory chains requires cofactors such as heme and copper and chaperones involved in the biogenesis of the enzyme complexes. The Gram-positive soil bacterium $\textit{Corynebacterium glutamicum}$ possesses a branched aerobic respiratory chain comprising, besides several dehydrogenases reducing menaquinone to menaquinol, a copper-dependent cytochrome $\textit{bc$_{1}$-aa$_{3}$}$ supercomplex and cytochrome $\textit{bd}$ quinol oxidase. The cytochrome $\textit{bc$_{1}$-aa$_{3}$}$ supercomplex is characteristic for aerobic $\textit{Actinobacteria}$ and the major generator of proton-motive force, but knowledge on its assembly or regulation is very limited. In this thesis, assembly factors involved in copper and heme insertion and regulatory proteins involved in expression control of respiratory chain components were identified and characterized. The following results were obtained: (i) Identification of the copper-deprivation stimulon led to the discovery of the two proteins Cg2699 ($\textbf{c}$opper $\textbf{t}$ransport and $\textit{i}$nsertion $\textbf{p}$rotein, CtiP) and Cg1884 (CopC). CtiP contains 16 predicted transmembrane helices and shows sequence similarity to the copper-transporter CopD and the cytochrome biogenesis chaperone CtaG. Deletion of $\textit{ctiP}$ resulted in a strong growth defect in standard glucose minimal medium (CGXII) resembling a cytochrome $\textit{aa$_{3}$}$ oxidase-deficient strain. Furthermore, the Δ$\textit{ctiP}$ strain exhibited an increased copper-tolerance, suggesting a copper transporting function. Transcriptome analysis revealed an induction of the copper-deprivation stimulon in the Δ$\textit{ctiP}$ 4 strain under copper sufficiency. CopC is a secreted protein with a C-terminal transmembrane helix and harbors a Cu(II)-binding site. Deletion of $\textit{copC}$ resulted in a growth defect in BHI complex medium and improved growth under copper excess, also suggesting an involvement in copper-transport. The lack of either CtiP or CopC prevented co-purification of the subunits of the supercomplex, indicating a crucial role of both CtiP and CopC in the correct assembly of the supercomplex. [...]


Note: Biotechnologie 1
Note: Dissertation, Heinrich-Heine-Universität Düsseldorf, 2019

Contributing Institute(s):
  1. Biotechnologie (IBG-1)
Research Program(s):
  1. 581 - Biotechnology (POF3-581) (POF3-581)

Appears in the scientific report 2019
Database coverage:
Creative Commons Attribution CC BY 4.0 ; OpenAccess
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Document types > Theses > Ph.D. Theses
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 Record created 2019-05-28, last modified 2022-09-30