Project

A balanced pool of deoxyribonucleotide triphosphates (dNTPs) is crucial for achieving successful nuclear DNA replication and repair. Temporal, spatial and ratio imbalances of intracellular dNTPs have been shown to have a mutagenic and cytotoxic effect to the cells. Therefore, it is essential to maintain the balance between dNTP biosynthesis and the degradation processes for cellular homeostasis. Multiple oncogenic signalling pathways feed into dNTP metabolism, and their imbalances play a conspicuous role in cancer initiation and progression. Balanced pools of dNTPs are essential for DNA replication to happen with the highest fidelity. Biased dNTP levels occurring at the replication site are mutagenic, and experimentally induced imbalances in dNTP pools have been associated with multiple mutations occurring randomly in the genome. In this project we seek to determine whether addition of thymidine (dThy) in cells, will increase the incidence of Ha-Ras Gly12 mutants. This event should be counteracted by addition of dCyd. By performing RNAseq, we expect to demonstrate the different events associated with an imbalanced dNTP pool leading to an increase in nuclear DNA repair mechanisms, the factors leading to the reduction of NTP to dNTP via the ribonucleotide reductase, the factors leading to apoptosis.