High Resolution cryo-EM Analysis of Ribosome-associated Functions

Objetivo

Translation of the genetically encoded information into polypeptides, protein biosynthesis, is a central function executed by ribosomes in all cells. In the case of membrane protein synthesis, integration into the membrane usually occurs co-translationally and requires a ribosome-associated translocon (SecYEG/Sec61). This highly coordinated process is poorly understood, since high-resolution structural information is lacking. Although single particle cryo-electron microscopy (cryo-EM) has given invaluable structural insights for such dynamic ribosomal complexes, the resolution is so far limited to 5-10 Å for asymmetrical particles. Thus, the mechanistic depth and reliability of interpretation has accordingly been limited.
Here, I propose to use single particle cryo-EM at improved, molecular resolution of 3-4 Å to study two fundamental ribosome-associated processes:
(i) co-translational integration of polytopic membrane proteins and
(ii) recycling of the eukaryotic ribosome.
First, we will visualize nascent polytopic membrane proteins inserting into the lipid bilayer via the bacterial ribosome-bound SecYEG translocon. Notably, the translocon will be embedded in a lipid environment provided by so-called nanodiscs. Second, we will visualize in a similar approach membrane protein insertion via the YidC insertase, the main alternative translocon. Third, as a novel research direction, we will determine the structure and function of eukaryotic ribosome recycling complexes involving the ABC-ATPase RLI.
The results will allow, together with functional biochemical data, an in-depth molecular structure-function analysis of these fundamental ribosome-associated processes. Moreover, reaching molecular resolution for asymmetrical particles by single particle cryo-EM will lift this technology to a level of analytical power approaching X-ray and NMR methods. ERC funding would allow for this highly challenging research to be conducted in an internationally competitive way in Europe.

Ámbito científico

Régimen de financiación

ERC-AG - ERC Advanced Grant

Institución de acogida

LUDWIG-MAXIMILIANS-UNIVERSITAET MUENCHEN

Aportación de la UE

€ 2 995 640,00

Dirección

GESCHWISTER SCHOLL PLATZ 1
80539 Muenchen
Alemania

Región

Bayern Oberbayern München, Kreisfreie Stadt

Tipo de actividad

Higher or Secondary Education Establishments

Investigador principal

Roland Beckmann (Prof.)

Contacto administrativo

Brice Rousseau (Mr.)

Enlaces

Contactar con la organización Sitio web

Coste total

Sin datos

Beneficiarios (1)