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TitleStructural basis for hygromycin B inhibition of yeast pseudouridine-deficient ribosomes.
Journal, issue, pagesSci Adv, Vol. 11, Issue 14, Page eadu0151, Year 2025
Publish dateApr 4, 2025
AuthorsYu Zhao / Chong Xu / Xin Chen / Hong Jin / Hong Li /
PubMed AbstractEukaryotic ribosomes are enriched with pseudouridine, particularly at the functional centers targeted by antibiotics. Here, we investigated the roles of pseudouridine in aminoglycoside-mediated ...Eukaryotic ribosomes are enriched with pseudouridine, particularly at the functional centers targeted by antibiotics. Here, we investigated the roles of pseudouridine in aminoglycoside-mediated translation inhibition by comparing the structural and functional properties of the yeast wild-type and the pseudouridine-free ribosomes. We showed that the pseudouridine-free ribosomes have decreased thermostability and high sensitivity to aminoglycosides. When presented with a model internal ribosomal entry site RNA, elongation factor eEF2, GTP (guanosine triphosphate), and sordarin, hygromycin B preferentially binds to the pseudouridine-free ribosomes during initiation by blocking eEF2 binding, stalling ribosomes in a nonrotated conformation. The structures captured hygromycin B bound at the intersubunit bridge B2a enriched with pseudouridine and a deformed codon-anticodon duplex, revealing a functional link between pseudouridine and aminoglycoside inhibition. Our results suggest that pseudouridine enhances both thermostability and conformational fitness of the ribosomes, thereby influencing their susceptibility to aminoglycosides.
External linksSci Adv / PubMed:40173234 / PubMed Central
MethodsEM (single particle)
Resolution2.38 - 2.49 Å
Structure data

EMDB-47093, PDB-9dov:
Hypopseudouridylated yeast 80S bound with Taura syndrome virus (TSV) internal ribosome entry site (IRES) and hygromycin B
Method: EM (single particle) / Resolution: 2.49 Å

EMDB-47098, PDB-9dp7:
Hypopseudouridylated yeast 80S bound with Taura syndrome virus (TSV) internal ribosome entry site (IRES) and hygromycin B, Class II
Method: EM (single particle) / Resolution: 2.38 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-HYG:
HYGROMYCIN B / antibiotic*YM

ChemComp-ZN:
Unknown entry

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • taura syndrome virus
KeywordsRIBOSOME / rRNA pseudouridylation / IRES initiation / confomation / eEF2 / hygromycin B

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