Mechanistic studies of prokaryotic genome defense mechanisms

Informazioni relative al progetto

MSOPGDM

ID dell’accordo di sovvenzione: 845268

DOI

10.3030/845268

Progetto chiuso

Data della firma CE 18 Marzo 2019

Data di avvio 1 Aprile 2019

Data di completamento 31 Marzo 2021

Finanziato da

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Costo totale

€ 191 149,44

Contributo UE

€ 191 149,44

Coordinato da

UNIVERSITAT ZURICH
Switzerland

Periodic Reporting for period 1 - MSOPGDM (Mechanistic studies of prokaryotic genome defense mechanisms)

Periodo di rendicontazione: 2019-04-01 al 2021-03-31

Sintesi del contesto e degli obiettivi generali del progetto

Prokaryotes are constantly threatened by a plethora of viruses (bacteriophages) and other genetic invaders. The evolutionary pressure associated with this host-parasite conflict has resulted in the emergence of numerous defense mechanisms that act on various stages of the bacteriophage life cycle. These defense mechanisms include restriction-modification enzymes and CRISPR-Cas systems, which have been proven to be useful tools for genome engineering. The current project set out to provide a mechanistic basis for immunity of two novel and uncharacterized prokaryotic defense systems. To address this question, we aimed to determine the molecular structures and biochemical activities of key effector proteins that are encoded by these systems. Deciphering these mechanisms will be important for our fundamental understanding of these systems that shape host-invader interactions and may additionally provide new tools to expand the genome engineering toolbox

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

The current project set out to provide a mechanistic basis for prokaryotic host immunity provided by two novel and uncharacterized prokaryotic defense systems. To address this question, we aimed to determine the molecular structures and biochemical activities of key effector proteins that are encoded by these systems. To this end, we established expression and purification protocols for all of the proteins of these systems and the respective proteins were isolated in sufficient quantities and qualities for structural and biochemical characterization. We succeeded in the determination of high-resolution molecular structures of key effector proteins using both X-ray crystallography and cryo-electron microscopy. Lastly, guided by the high-resolution structures we obtained the first insights into the biochemical activities of the systems. The results obtained within the timeframe of this proposal provide a foundation for future studies that advance our understanding the biological functions and mechanisms of prokaryotic host defense systems.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

Despite setbacks due to the ongoing covid-19 pandemic, the goals of the project have been partially reached, as the studies conducted in the course of the project have yielded publishable results, which are currently being prepared for publication. The results will also provide a foundation for future mechanistic and functional studies of prokaryotic defense systems.

Mechanistic studies of prokaryotic genome defense mechanisms