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TitleAlternative Mode of E-Site tRNA Binding in the Presence of a Downstream mRNA Stem Loop at the Entrance Channel.
Journal, issue, pagesStructure, Vol. 26, Issue 3, Page 437-445.e3, Year 2018
Publish dateMar 6, 2018
AuthorsYan Zhang / Samuel Hong / Ajchareeya Ruangprasert / Georgios Skiniotis / Christine M Dunham /
PubMed AbstractStructured mRNAs positioned downstream of the ribosomal decoding center alter gene expression by slowing protein synthesis. Here, we solved the cryo-EM structure of the bacterial ribosome bound to an ...Structured mRNAs positioned downstream of the ribosomal decoding center alter gene expression by slowing protein synthesis. Here, we solved the cryo-EM structure of the bacterial ribosome bound to an mRNA containing a 3' stem loop that regulates translation. Unexpectedly, the E-site tRNA adopts two distinct orientations. In the first structure, normal interactions with the 50S and 30S E site are observed. However, in the second structure, although the E-site tRNA makes normal interactions with the 50S E site, its anticodon stem loop moves ∼54 Å away from the 30S E site to interact with the 30S head domain and 50S uL5. This position of the E-site tRNA causes the uL1 stalk to adopt a more open conformation that likely represents an intermediate state during E-site tRNA dissociation. These results suggest that structured mRNAs at the entrance channel restrict 30S subunit movement required during translation to slow E-site tRNA dissociation.
External linksStructure / PubMed:29456023 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 3.5 Å
Structure data

8596

5uq7

EMDB-8596, PDB-5uq7:
70S ribosome complex with dnaX mRNA stemloop and E-site tRNA ("in" conformation)
Method: EM (single particle) / Resolution: 3.5 Å

8597

5uq8

EMDB-8597, PDB-5uq8:
70S ribosome complex with dnaX mRNA stem-loop and E-site tRNA ("out" conformation)
Method: EM (single particle) / Resolution: 3.2 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-SF4:
IRON/SULFUR CLUSTER / Iron–sulfur cluster

Source
  • thermus thermophilus (strain hb8 / atcc 27634 / dsm 579) (bacteria)
  • escherichia coli k-12 (bacteria)
  • synthetic construct (others)
KeywordsRIBOSOME / 70S ribosome / dnaX / stem-loop / structured mRNA / -1 programmed frameshifting / E site

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