DNA damage can arise from both internal (ROS, replication stress) and external (UV, IR, chemotherapeutic agents) sources. Upon DNA damage, signaling and repair pathways are activated that are important for cell survival. As DNA damage can lead to cancer and is used in cancer treatment, understanding the basic mechanisms of DNA damage repair may help us both to prevent cancer and make cancer treatment more efficient in the future.
How the DNA damage signaling and repair pathways interact with the cellular transcription machinery is currently a topic of high interest in the field. During the transcription cycle, RNA polymerase II (RNAPII), which transcribes DNA into RNA, becomes phosphorylated in its C-terminal domain (CTD). By studying the RNAPII CTD phosphatase complex WDR82/PNUTS-PP1, we recently showed that when phosphorylated, the RNAPII CTD promotes DNA damage signaling and conflicts between transcription and replication. Our main hypothesis is thus that the RNAPII CTD may serve as a central signaling platform in DNA damage signaling and repair pathways. However, whether the RNAPII CTD promotes repair of DNA double strand breaks, the most lethal type of DNA damage, is currently not known.
Project:
Our hypothesis is that the RNAPII CTD promotes repair of DNA double strand breaks via the non-homologous end joining pathway of DNA double strand break repair. This is currently an ongoing project in the group, and the master student project will be incorporated into this. To address our hypothesis, we will use a variety of molecular biology techniques, such as DNA repair assays, protein- protein interaction studies and siRNA mediated depletion and rescue experiments. The results will contribute with highly novel knowledge into the mechanisms of DNA damage repair.
Methods:
The student will learn techniques that are frequently used in our group, such as cell culture and transfection, western blotting, protein pulldowns and multiparameter flow cytometry including barcoding. He or she will work closely with senior scientist Helga Landsverk (who will also be the main supervisor) in the group ‘Radiation Biology and DNA Damage Signaling’ at the Department of Radiation Biology, Institute for Cancer Research. The group leader, Randi Sylju?sen, will be co-supervisor on this project.
The project is suitable for a dedicated student who wants:
-- extensive hands-on experience in molecular biology techniques
-- to contribute to novel discoveries about basic molecular mechanisms
-- a project in the field of molecular cancer research