Logo: to the web site of Uppsala University

uu.sePublikasjoner fra Uppsala universitet
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Insights into Translocation Mechanism and Ribosome Evolution from Cryo-EM Structures of Translocation Intermediates of Giardia intestinalis
Vise andre og tillknytning
(engelsk)Manuskript (preprint) (Annet vitenskapelig)
Abstract [en]

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 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 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 both from the structural and mechanistic perspectives.

HSV kategori
Identifikatorer
URN: urn:nbn:se:uu:diva-495331OAI: oai:DiVA.org:uu-495331DiVA, id: diva2:1731150
Tilgjengelig fra: 2023-01-26 Laget: 2023-01-26 Sist oppdatert: 2023-01-26
Inngår i avhandling
1. Cryo-EM and Computational Biology of Macromolecular Complexes
Åpne denne publikasjonen i ny fane eller vindu >>Cryo-EM and Computational Biology of Macromolecular Complexes
2023 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

The ribosome is a large, ancient multicomponent macromolecular complex which is highly amenable to study by cryogenic electron microscopy (cryo-EM) and computation biology methods. This thesis delves into the structure of both prokaryotic and eukaryotic ribosomes in the context of determining a solution to emerging antimicrobial resistance. We show that thermorubin (THB) binds to the E. coli ribosome at intersubunit bridge B2a, flipping out 23S rRNA residue C1914 which interferes with A-site substrates. The position and rearrangements caused by THB also accounts for the biochemical results showing a decrease in elongation, termination and recycling phases of translation. Also using cryo-EM we looked at the Giardia intestinalis ribosome, determining six high-resolution structures representing translocation intermediates. Giardia is a protozoan parasite causing diarrhoea in humans, with metronidazole strains emerging. As the ribosome is often a target for antimicrobial drugs, work on the structure and function of the ribosome is of utmost important in determining an alternative therapeutic approach to the treatment of giardiasis. We also show naturally bound tRNAs and eEF2 on the Giardia ribosome, exhibiting eukaryote-specific subunit rolling and eEF2 with GDP in a uniquely positioned Pi primed for release, adding to the mechanism of translocation in protists as well as illustrating the evolution of both the structure and function of translation machinery. Finally, the molecular basis of thermostability in translational GTPases is explored using molecular dynamics of mesophilic and thermophilic elongation factor EF-Tu. Through ancestral sequence reconstruction two key interactions: in the GTPase domain; and an interdomain interaction were shown to be important in the overall structural stability of EF-Tu in high temperature environments. These studies together highlight the strength of utilising both structural and computational techniques to explore the translation apparatus.

sted, utgiver, år, opplag, sider
Uppsala: Acta Universitatis Upsaliensis, 2023. s. 45
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 2232
HSV kategori
Forskningsprogram
Biologi med inriktning mot molekylärbiologi
Identifikatorer
urn:nbn:se:uu:diva-495335 (URN)978-91-513-1698-7 (ISBN)
Disputas
2023-03-17, A1:107a, Biomedicinskt centrum, Husargatan 3, Uppsala, 09:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2023-02-24 Laget: 2023-01-26 Sist oppdatert: 2023-02-24

Open Access i DiVA

Fulltekst mangler i DiVA

Søk utenfor DiVA

GoogleGoogle Scholar

urn-nbn

Altmetric

urn-nbn
Totalt: 252 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf