Antibiotic thermorubin tethers ribosomal subunits and impedes A-site interactions to perturb protein synthesis in bacteria

Parajuli, Narayan Prasad,1989-Uppsala universitet,Institutionen för cell- och molekylärbiologi (author)

Thermorubin (THB) is a long-known broad-spectrum ribosome-targeting antibiotic, but the molecular mechanism of its action was unclear. Here, our precise fast-kinetics assays in a reconstituted Escherichia coli translation system and 1.96 Å resolution cryo-EM structure of THB-bound 70S ribosome with mRNA and initiator tRNA, independently suggest that THB binding at the intersubunit bridge B2a near decoding center of the ribosome interferes with the binding of A-site substrates aminoacyl-tRNAs and class-I release factors, thereby inhibiting elongation and termination steps of bacterial translation. Furthermore, THB acts as an anti-dissociation agent that tethers the ribosomal subunits and blocks ribosome recycling, subsequently reducing the pool of active ribosomes. Our results show that THB does not inhibit translation initiation as proposed earlier and provide a complete mechanism of how THB perturbs bacterial protein synthesis. This in-depth characterization will hopefully spur efforts toward the design of THB analogs with improved solubility and effectivity against multidrug-resistant bacteria.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
NATURVETENSKAP Biologi Strukturbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Structural Biology hsv//eng
Antibiotic
Thermorubin
Ribosome
Translation inhibition
Subunit tethering
Biokemi
Biochemistry
Biology with specialization in Molecular Biology
Biologi med inriktning mot molekylärbiologi
Biology with specialization in Structural Biology
Biologi med inriktning mot strukturbiologi

Emmerich, Andrew GUppsala universitet,Institutionen för cell- och molekylärbiologi(Swepub:uu)andem552 (author)
Mandava, Chandra Sekhar,1978-Uppsala universitet,Institutionen för cell- och molekylärbiologi(Swepub:uu)chsma839 (author)
Pavlov, Michael Y.Uppsala universitet,Institutionen för cell- och molekylärbiologi(Swepub:uu)mpa27542 (author)
Sanyal, SuparnaUppsala universitet,Institutionen för cell- och molekylärbiologi(Swepub:uu)sucsa102 (author)
Uppsala universitetInstitutionen för cell- och molekylärbiologi (creator_code:org_t)

By the author/editor: Parajuli, Naraya ...; Emmerich, Andrew ...; Mandava, Chandra ...; Pavlov, Michael ...; Sanyal, Suparna

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Structural Biolo ...

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