Puck Knipscheer Group
Molecular mechanisms and regulation of DNA repair
The main interest of our laboratory is to decipher the molecular details of cellular processes that maintain the integrity of our genome. Currently, our work focuses on two such processes.
- First, the repair of highly toxic DNA interstrand crosslinks (ICLs). These lesions can form endogenously but are also induced at high doses in cancer chemotherapy. Repair of these lesions requires several DNA repair pathways including the Fanconi anemia (FA) pathway but the molecular mechanism is poorly understood.
- Second, we aim to understand how stable secondary DNA structures (for example G4 structures) are resolved. G4 structures have various cellular functions but can also cause genome instability under certain cellular conditions. For example, G4 sequences are found specifically enriched at chromosomal breakpoints in human cancers. To understand how these stable DNA structures can cause mutations it is important to understand the mechanisms that resolve these structures.
We study these processes in the context of active DNA replication and make use of Xenopus egg extracts, a unique system that supports highly regulated vertebrate DNA replication in vitro. In this experimental system we can recapitulate both ICL repair and G4 structure replication in a test tube, allowing us to gain insights in the molecular mechanisms. We combine these studies with mass spectrometry and biochemical, biophysical and cell biological techniques to obtain a detailed and complete understanding of these processes that guard our genome.