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Structure paper

TitlemRNA stem-loops can pause the ribosome by hindering A-site tRNA binding.
Journal, issue, pagesElife, Vol. 9, Year 2020
Publish dateMay 19, 2020
AuthorsChen Bao / Sarah Loerch / Clarence Ling / Andrei A Korostelev / Nikolaus Grigorieff / Dmitri N Ermolenko /
PubMed AbstractAlthough the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via ...Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from mRNA and the transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation.
External linksElife / PubMed:32427100 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 3.4 Å
Structure data

21420

6vwl

EMDB-21420, PDB-6vwl:
70S ribosome bound to HIV frameshifting stem-loop (FSS) and P/E tRNA (rotated conformation)
Method: EM (single particle) / Resolution: 3.1 Å

21421

6vwm

EMDB-21421, PDB-6vwm:
70S ribosome bound to HIV frameshifting stem-loop (FSS) and P-site tRNA (non-rotated conformation, Structure I)
Method: EM (single particle) / Resolution: 3.4 Å

21422

6vwn

EMDB-21422, PDB-6vwn:
70S ribosome bound to HIV frameshifting stem-loop (FSS) and P-site tRNA (non-rotated conformation, Structure II)
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • Eschericia coli (E. coli)
  • escherichia coli (E. coli)
  • human immunodeficiency virus 1
  • Homo sapiens (human)
KeywordsRIBOSOME / HIV FSS / frameshifting

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