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• W4366334211 abstract "Abstract Ribosome hibernation is a key survival strategy bacteria adopt under environmental stress, where a protein, hibernation promotion factor (HPF), transitorily inactivates the ribosome and slows down its overall protein synthesis. The mechanism is well studied in enteric bacteria, which mainly hibernate its ribosome in 100S disome form through a dual domain, long HPF (HPF long ) or a single domain, short HPF (HPF short ) in concert with another ribosome modulation factor. Mycobacteria under hypoxia (low oxygen) stress overexpresses RafH protein regulated under DosR regulon, a critical factor for its survival. The RafH, a dual domain HPF, an orthologue of bacterial HPF long , hibernates ribosome in 70S monosome form only. Here we report the cryo-EM structure of Mycobacterium smegmatis , a close homologue of M. tuberculosis , 70S ribosome in complex with the RafH factor at an overall 2.8 Å resolution. The RafH N-terminus domain (NTD) is conserved and binds to the decoding center of the ribosomal small subunit, a similar binding site of HPF long NTD, but additionally it also interacts with the inter subunit bridge, B2a. Contrary to the HPF long CTD, the RafH CTD, which is larger, binds to a unique site at the platform binding center of the ribosomal small subunit and sandwiches between bS1 and uS11 ribosomal proteins. The two domain connecting linker regions, which remain mostly disordered in earlier reported HPF long structures, interacts mainly with the anti-Shine Dalgarno sequence of the 16S rRNA. The helix H54a of 23S rRNA, unique to the mycobacterial ribosome, adopts a different conformation and come close to RafH CTD, suggesting its role in ribosome hibernation. RafH inhibits in-vitro protein synthesis in a concentration dependent manner. Further, the modeling studies provided the structural basis for the incompatibility of mycobacterial ribosomes forming 100S like hibernating ribosomes." @default.
• W4366334211 created "2023-04-20" @default.
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• W4366334211 date "2023-04-18" @default.
• W4366334211 modified "2023-09-26" @default.
• W4366334211 title "Cryo-EM structure of the mycobacterial 70S ribosome in complex with ribosome hibernation promotion factor RafH, reveals the unique mode of mycobacterial ribosome hibernation" @default.
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• W4366334211 doi "https://doi.org/10.1101/2023.04.18.537051" @default.
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