H2020Индивидуална стипендия2019–2021

RespViRALI · Structural and functional insights into the assembly of respiratory complexes by a novel putative chaperone

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2019-07-01 → 2021-06-30
Финансиране от ЕС: 173 076 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Протеините ViaA, RavA и LdcI в бактерията E. coli се изследват като помощни механизми за сглобяване на дихателните комплекси Nuo и Frd. Разбирането на тези процеси помага да се обясни устойчивостта на бактериите към определени антибиотици и развитието на невродегенеративни заболявания.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Structural and functional insights into the assembly of respiratory complexes by a novel putative chaperone

Cellular respiratory complexes play a central role in the production of ATP, the universal energy source of life. Defects in their activity are linked to neurodegenerative diseases in humans and antibiotics resistance in bacteria. Despite their capital importance, the assembly mechanisms of respiratory complexes remain highly unexplored. The goal of this project was to investigate how the assembly of the major bacterial respiratory Complex I is assisted by a multi-protein system involved in enterobacterial stress response. More specifically, I explored the potential role of E. coli proteins ViaA, RavA and LdcI as a macromolecular machine assisting assembly of bacterial respiratory Complex I (Nuo) and fumarate reductase (Frd). RavA is an AAA+ ATPase of the MoxR type, a protein family of which the members are suggested to be involved in chaperone functions. RavA is involved in the bacterial acid-stress response by modulating the activity of the acid-stress inducible lysine decarboxylase (LcdI) during stringent response, and is shown to interact with ViaA, an operon partner of RavA containing a Von-Willebrand Factor A (VWA) domain. The rationale for the involvement of LdcI, RavA and ViaA in the assembly of Nuo and Frd is that : 1) RavA and LdcI together form a huge cage-like assembly which is shown to interact with specific subunits of Nuo and Frd. 2) ViaA, a protein of unknown structure proposed to act as a shuttle protein between the LdcI-RavA cage and its substrates, was also shown to interact with Nuo and Frd subunits. 3) E. coli cells lacking ravA/viaA genes acquire resistance to aminoglycosides, a class of antibiotics requiring proton motive force, generated by bacterial respiratory complex 1, for their uptake. We aimed to elucidate the involvement of the LdcI-RavA-ViaA triad in the assembly and maturation of Nuo and Frd by a combination of biochemical/biophysical methods to validate interactions between LdcI, RavA and ViaA and specific soluble sub-complexes of Nuo and Frd, with a structural characterization of the most promising interactions using an integrated structural biology approach combining low- and high-resolution techniques.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Cellular respiratory complexes play a central role in the production of ATP, the universal energy source of life. Defects in their activity are linked to neurodegenerative diseases in humans and antibiotics resistance in bacteria. Despite their capital importance, the assembly mechanisms of respiratory complexes remain highly unexplored.This research project aims to investigate how the assembly of the major bacterial respiratory Complex I is assisted by a multi protein system involved in enterobacterial stress response. The host team recently solved the structures of two components of this system with cryo-electron microscopy (cryoEM) and X-ray crystallography, and showed that they form a huge cage-like assembly reminiscent of the GroEL-ES chaperone. Importantly, the latest scientific literature reported physical interactions between this macromolecular assembly and specific subunits of E. coli respiratory complexes.To elucidate the functional role of this enigmatic machine in the assembly of Complex I, I will employ a highly integrated approach situated at the front line of molecular structural biology. A wide range of biophysical methods will be used to dissect the affinity and kinetics of individual interactions between components of this multi protein system and subunits of Complex I. I will examine the overall architecture of the formed complexes with negative stain electron microscopy and small-angle X-Ray/Neutron scattering, while atomic resolution insights of the complexes will be provided by high resolution single particle cryoEM and X-ray crystallography.The structures and functional insights obtained in the proposed project will improve our general understanding of coordination between protein folding and assembly of respiratory complexes, and bring novel insights into their implication in antibiotics resistance and pathways of bacterial stress responses, which are crucial to developing strategies for controlling infection.

Оригинален текст от CORDIS (на английски).

Участници

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS · ParisКоординаторФранция