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This lecture by Eilika Weber-Ban from ETH Zürich explores the molecular machines involved in mycobacterial protein quality control pathways, focusing on the degradation pathways in protein homeostasis, the compartmentalizing proteases in Mycobacterium tuberculosis, and the pupylation-dependent proteasomal degradation pathway. The lecture delves into the recognition of Clp substrate classes, the asymmetric nature of Mycobacterial ClpP, and the role of Pup as a recruitment tag and threading initiator. It also discusses the importance of Mpa-proteasome complex in substrate selection, the depupylation rates of different substrates, and the mechanism and evolutionary origin of pupylation enzymes.