A biochemical and genetic characterization of DNA polymerase epsilon to understand its role in DNA repair, cell cycle regulation, and DNA replication

Project description

The diploid human genome has been estimated to contain 6 billion nucleotides. It is an enormous task for the growing cell to replicate the entire genome, due to the accuracy by which it must be accomplished. It must also be a very efficient process because the cell has a very limited time to finish the task. We are studying on the molecular level how DNA replication is accomplished in eukaryotic cells. There still remain many large questions that have not been resolved. It is not known how DNA replication is initiated, or how a replication fork is functioning. We are focused on the proteins that participate at the eukaryotic replication fork. Our main focus lies on DNA polymerase epsilon that may play a role at the replication fork, although its precise function has not been determined.

Our maingoals are,

(i) to understand how DNA polymerase epsilon interacts with other replication fork proteins,

(ii) to understand whether DNA polymerase epsilon participates at the replication fork or if DNA polymerase epsilon fulfills another important function during the replication of the chromosomes,

(iii) to understand how DNA polymerase epsilon contributes to the fidelity of the replication of the genome,

(iv) to understand how DNA polymerase epsilon is involved in DNA repair processes and cell-cycle regulation.

Our model system is S. cerevisiae and this allows us to complement our biochemical characterizations with yeast genetics in in vivo experiments.