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The assembly of the membrane arm of mitochondrial complex I follows evolutionarily distinct routes

Parlar, S.; Marofke, M.; Kühn, K.; Gawryluk, R.; Meyer, E.

2025-06-11 biochemistry
10.1101/2025.06.10.658862 bioRxiv
Show abstract

Complex I is the largest enzyme of the mitochondrial oxidative phosphorylation system. It is composed of core subunits that are conserved across bacteria and eukaryotes and accessory subunits that are lineage-specific. Consequently, the assembly of complex I likely requires lineage-specific assembly machineries. Complex I assembly has been mainly studied in mammals. It follows a modular pathway in which modules are assembled independently and then associated together to form the mature complex. Here, we identify and characterize PAF1 as an assembly factor for the membrane arm of complex I in plants. We show that this protein plays a role in stabilizing a module of the membrane arm prior to its association with the rest of the complex. PAF1 was likely present in the last eukaryotic common ancestor and has been lost in mammals in which the assembly of the membrane arm involves assembly factors not conserved in plants. Altogether, we provide molecular evidence that the assembly of the membrane arm of complex I can be performed by two different machineries.

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