Mechanistic studies of prokaryotic genome defense mechanisms

Información del proyecto

MSOPGDM

Identificador del acuerdo de subvención: 845268

DOI

10.3030/845268

Proyecto cerrado

Fecha de la firma de la CE 18 Marzo 2019

Fecha de inicio 1 Abril 2019

Fecha de finalización 31 Marzo 2021

Financiado con arreglo a

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coste total

€ 191 149,44

Aportación de la UE

€ 191 149,44

Coordinado por

UNIVERSITAT ZURICH
Switzerland

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

Período documentado: 2019-04-01 hasta 2021-03-31

Resumen del contexto y de los objetivos generales del proyecto

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

Trabajo realizado desde el comienzo del proyecto hasta el final del período abarcado por el informe y los principales resultados hasta la fecha

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.

Avances que van más allá del estado de la técnica e impacto potencial esperado (incluida la repercusión socioeconómica y las implicaciones sociales más amplias del proyecto hasta la fecha)

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