Descrizione del progetto DEENESFRITPL Decodificare il co-co-assemblaggio Un assemblaggio di componenti combinati assieme in fasi diverse conduce a un co-co-assemblaggio. Inoltre, il co-co assemblaggio è impiegato principalmente per la formazione di omo-oligomeri ed è presente in tutti i regni degli esseri viventi. Tuttavia, non si sa ancora molto circa i meccanismi molecolari alla sua base. Il progetto Co-coAssembly, finanziato dall’UE, studierà i meccanismi di co-co-assemblaggio utilizzando la corismato mutasi dimerica come principale candidato rappresentativo da uno schermo ad alte prestazioni per il co-co-assemblaggio di complessi proteici nell’E. coli. Attraverso una tomografia speciale, lo studio si concentrerà sull’organizzazione tridimensionale dei ribosomi di E. coli per valutare il modo in cui l’organizzazione di macchinari traslazionali consente il co-co-assemblaggio. Mostra l’obiettivo del progetto Nascondi l’obiettivo del progetto Obiettivo Almost all fundamental biological processes involve protein complexes and therefore, efficient folding and assembly of homo- and hetero-oligomers is critical for cellular functionality and integrity. Recent studies have shown that many protein complexes assemble co-translationally by one fully-synthesized subunit engaging another subunit that is still in nascent state (co-post assembly). An ongoing study in the Bukau lab now revealed that assembly can also occur by interaction of two partner nascent chains (co-co assembly). Co-co assembly is mostly employed for the formation of homo-oligomers and exists in all kingdoms of life.. Despite initial evidence of its existence, very little is known about the molecular mechanisms driving co-co assembly. This includes information on whether co-co assembly requires co-localization of two polysomes or can happen on one polysome.. Furthermore, it is currently unclear whether co-co interactions require preceding nascent chain folding steps and to what extent co-translationally acting chaperones coordinate the process and the impact of translation speed on co-co assembly.I propose to study mechanisms of co-co assembly using the dimeric chorismate mutase (PheA) as a representative top candidate from a high throughput screen for co-co assembling protein complexes in E. coli. Employing cryo-electron tomography, I will analyse the three dimensional arrangement of E. coli ribosomes in the context of a polysome to assess how organization of translational machinery allows co-co assembly. Moreover, I will study the co-translational cascade of folding steps of chorismate mutase by utilizing FRET on in vitro prepared nascent chains. Finally, I plan to explore the impact of co-translationally acting chaperones and translation kinetics on co-co assembly, by performing disome-selective profiling analysis in chaperone mutant cells lacking Trigger Factor and DnaK and in mutants that synthesize proteins with reduced translation kinetics. Campo scientifico scienze naturaliscienze biologichebiochimicabiomolecoleproteinescienze naturaliscienze fisicheotticamicroscopiaelectron microscopy Programma(i) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Argomento(i) MSCA-IF-2019 - Individual Fellowships Invito a presentare proposte H2020-MSCA-IF-2019 Vedi altri progetti per questo bando Meccanismo di finanziamento MSCA-IF-EF-RI - RI – Reintegration panel Coordinatore RUPRECHT-KARLS-UNIVERSITAET HEIDELBERG Contribution nette de l'UE € 174 806,40 Indirizzo Seminarstrasse 2 69117 Heidelberg Germania Mostra sulla mappa Regione Baden-Württemberg Karlsruhe Heidelberg, Stadtkreis Tipo di attività Higher or Secondary Education Establishments Collegamenti Contatta l’organizzazione Opens in new window Sito web Opens in new window Partecipazione a programmi di R&I dell'UE Opens in new window Rete di collaborazione HORIZON Opens in new window Altri finanziamenti € 0,00
