Unravelling the mechanism of eukaryotic helicase activation

Información del proyecto

MechHelicaseActiv8on

Identificador del acuerdo de subvención: 101025825

DOI

10.3030/101025825

Proyecto cerrado

Fecha de la firma de la CE 27 Abril 2021

Fecha de inicio 1 Septiembre 2022

Fecha de finalización 31 Agosto 2024

Financiado con arreglo a

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coste total

€ 212 933,76

Aportación de la UE

€ 212 933,76

Coordinado por

THE FRANCIS CRICK INSTITUTE LIMITED
United Kingdom

Periodic Reporting for period 1 - MechHelicaseActiv8on (Unravelling the mechanism of eukaryotic helicase activation)

Período documentado: 2022-09-01 hasta 2024-08-31

Resumen del contexto y de los objetivos generales del proyecto

The project aimed to unravel the mechanism of eukaryotic helicase activation in DNA replication initiation. During DNA replication, helicase is essential for unwinding the DNA double helix, a step critical for cellular division. However, the exact mechanisms by which the helicase is activated to start DNA unwinding, including the transition from double-stranded to single-stranded DNA, remain unclear.
Understanding DNA replication mechanisms is fundamental to advancing knowledge in genome maintenance. Errors in DNA replication can lead to genetic mutations, contributing to cancer and other genetic disorders. By shedding light on helicase activation, this research has the potential to aid in the development of targeted therapies for diseases that result from DNA replication errors.
The main objectives of the project were: (1) to uncover the fundamental mechanisms by which helicase is activated to initiate DNA unwinding, (2) to investigate the specific roles of various protein components within the helicase complex.

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

For helicase to get activated during DNA replication initiation, the helicase has to transit from double stranded DNA to single stranded DNA. To find out the trajectory of lagging strand template during helicase activation, I purified all essential DNA replication proteins and confirmed their activity in reconstituted DNA replication assay. I prepared a set of helicase mutants required for testing the helicase activation mechanism.
Mcm10 assists helicase activation by supporting the transition from double stranded DNA to single stranded DNA and increasing replication speed. Together with our collaborators, we combined biochemistry with structural biology to further understand the mechanism of Mcm10 action. We identified distinct roles for the N-terminal (initiates replication) and C-terminal (increases speed) of Mcm10. Mutations in Mcm10 confirmed that its interaction with single stranded DNA is not essential for replication. We disseminated the findings by publishing in Nature Structural Biology with Open Acces.

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)

The project contributed to our understanding of Mcm10 function as well as provided a separation-of-function mutant that might be useful for further research in the field. The helicase mutants might be further used in testing other processes, including how the helicase bypass DNA lesion. This could lead to further insight on not only the mechanism of DNA replication, but also other processes like DNA repair, which is important for genome stability.