An Archaea-specific -type cytochrome maturation machinery is crucial for methanogenesis in .

-Type cytochromes (cyt ) are proteins that undergo post-translational modification to covalently bind heme, which allows them to facilitate redox reactions in electron transport chains across all domains of life. Genomic evidence suggests that cyt are involved in electron transfer processes among the Archaea, especially in members that produce or consume the potent greenhouse gas methane. However, neither the maturation machinery for cyt in Archaea nor their role in methane metabolism has ever been functionally characterized. Here, we have used CRISPR-Cas9 genome editing tools to map a distinct pathway for cyt biogenesis in the model methanogenic archaeon and have also identified substrate-specific functional roles for cyt during methanogenesis. Although the cyt maturation machinery from is universally conserved in the Archaea, our evolutionary analyses indicate that different clades of Archaea acquired this machinery through multiple independent horizontal gene transfer events from different groups of Bacteria. Overall, we demonstrate the convergent evolution of a novel Archaea-specific cyt maturation machinery and its physiological role during methanogenesis, a process which contributes substantially to global methane emissions.