Investigating eukaryotic replisome dynamics at the single molecule level

Cel

For cells to reproduce, an accurate duplicate of the genome must be created. This is no small task. The genetic information stored in each cell consists of ~6 billion pairs of nucleobases (base pairs, bp) assembled as a polymer 2 metres long and 2 nanometres in diameter, with the structural form of a double helix. For a mammalian cell to divide, this deoxyribonucleic acid (DNA) must be copied in a time frame on the order of 1 day, or ~70,000bp a second. DNA replication is common to all 3 domains of life, bacteria, archaea and eukarya and is accomplished by a complex of proteins. This proposal brings together a researcher of great proficiency in single molecule methods and multidisciplinary research with the Single Molecule Imaging group at the London Research Institute, one of the world leading centres in DNA replication. Combined, we will build unique instruments and develop single molecule assays to understand the molecular gymnastics of DNA replication in eukaryotes. We will elucidate rates of DNA unwinding by eukaryotic helicases and establish enhancements by association with other proteins. We will also study replisome dynamics by observing synthesis of DNA on custom templates in real time. This will allow detection of replication loops and stalling that may occur. We will also examine the mechanism of lesion bypass. The insight gained is impossible with classical biochemical techniques, as individual replisomes are observed in real time rather than measuring an average of a population. Our methods will reveal heterogeneities and obtain precise quantitative details of the dynamics. Features such as pauses and back slips will enable the study of intermediate states and conformational changes linked to replisome dynamics. This proposal will satisfy academic curiosity of understanding life at the most fundamental level but will also increase our knowledge of the how the cell works and thus becomes the building blocks for disease treatment and cures of the future.

Dziedzina nauki

Program(-y)

Temat(-y)

MSCA-IF-2014-EF - Marie Skłodowska-Curie Individual Fellowships (IF-EF)

Zaproszenie do składania wniosków

H2020-MSCA-IF-2014

Zobacz inne projekty w ramach tego zaproszenia

System finansowania

MSCA-IF-EF-ST - Standard EF

Koordynator

THE FRANCIS CRICK INSTITUTE LIMITED

Wkład UE netto

€ 195 454,80

Adres

1 MIDLAND ROAD
NW1 1AT London
Zjednoczone Królestwo

Region

London Inner London — West Camden and City of London

Rodzaj działalności

Research Organisations

Linki

Kontakt z organizacją Strona internetowa

Uczestnictwo w unijnych programach w zakresie badań i innowacji

sieć współpracy HORIZON

Koszt całkowity

€ 195 454,80

Uczestnicy (1)

CANCER RESEARCH UK LBG

Zjednoczone Królestwo