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TitleInsights into translocation mechanism and ribosome evolution from cryo-EM structures of translocation intermediates of Giardia intestinalis.
Journal, issue, pagesNucleic Acids Res, Vol. 51, Issue 7, Page 3436-3451, Year 2023
Publish dateMar 13, 2023
AuthorsSoneya Majumdar / Andrew Emmerich / Sascha Krakovka / Chandra Sekhar Mandava / Staffan G Svärd / Suparna Sanyal /
PubMed AbstractGiardia intestinalis is a protozoan parasite that causes diarrhea in humans. Using single-particle cryo-electron microscopy, we have determined high-resolution structures of six naturally populated ...Giardia intestinalis is a protozoan parasite that causes diarrhea in humans. Using single-particle cryo-electron microscopy, we have determined high-resolution structures of six naturally populated translocation intermediates, from ribosomes isolated directly from actively growing Giardia cells. The highly compact and uniquely GC-rich Giardia ribosomes possess eukaryotic rRNAs and ribosomal proteins, but retain some bacterial features. The translocation intermediates, with naturally bound tRNAs and eukaryotic elongation factor 2 (eEF2), display characteristic ribosomal intersubunit rotation and small subunit's head swiveling-universal for translocation. In addition, we observe the eukaryote-specific 'subunit rolling' dynamics, albeit with limited features. Finally, the eEF2·GDP state features a uniquely positioned 'leaving phosphate (Pi)' that proposes hitherto unknown molecular events of Pi and eEF2 release from the ribosome at the final stage of translocation. In summary, our study elucidates the mechanism of translocation in the protists and illustrates evolution of the translation machinery from bacteria to eukaryotes from both the structural and mechanistic perspectives.
External linksNucleic Acids Res / PubMed:36912103 / PubMed Central
MethodsEM (single particle)
Resolution3.25 - 6.49 Å
Structure data

16211

8br8

EMDB-16211, PDB-8br8:
Giardia ribosome in POST-T state (A1)
Method: EM (single particle) / Resolution: 3.35 Å

16222

8brm

EMDB-16222, PDB-8brm:
Giardia ribosome in POST-T state, no E-site tRNA (A6)
Method: EM (single particle) / Resolution: 3.33 Å

16225

8bsi

EMDB-16225, PDB-8bsi:
Giardia ribosome chimeric hybrid-like GDP+Pi bound state (B1)
Method: EM (single particle) / Resolution: 3.4 Å

16226

8bsj

EMDB-16226, PDB-8bsj:
Giardia Ribosome in PRE-T Classical State (C)
Method: EM (single particle) / Resolution: 6.49 Å

16228

8btd

EMDB-16228, PDB-8btd:
Giardia Ribosome in PRE-T Hybrid State (D1)
Method: EM (single particle) / Resolution: 4.9 Å

16235

8btr

EMDB-16235, PDB-8btr:
Giardia Ribosome in PRE-T Hybrid State (D2)
Method: EM (single particle) / Resolution: 3.25 Å

Chemicals

ChemComp-PO4:
PHOSPHATE ION / Phosphate

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM / Guanosine diphosphate

Source
  • giardia lamblia atcc 50803 (eukaryote)
  • giardia intestinalis (eukaryote)
KeywordsRIBOSOME / Translation / Giardia / Eukaryote / Eukaryotic / Macromolecule / tRNA / EF-2 / Elongation / GDP

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