Understanding Bacterial Nucleotide Excision Repair at the Level of the Single Molecule Inside Living Cells.
Malfunctioning DNA repair lead to an accumulation of mutations, which frequently results in cancer. The Nucleotide Excision Repair (NER) pathway removes a DNA lesions caused by UV light, cigarette smoke and chemical mutagens. NER is highly conserved, and studying the simpler NER in bacteria provides key insight into human NER. I propose an interdisciplinary approach to understand the mechanistic details of bacterial NER in living cells. I will use a combination of cutting-edge single-molecule methods to elucidate how DNA is repaired inside living cells. I will use super-resolution microscopy combined to study the behaviour of individual NER proteins. To complement this, conventional biochemistry, cell biology, genetics, smFRET assays, FCS and TIRF microscopy will be used. Together, this will provide a comprehensive understanding of the bacterial NER pathway, and constitute the first steps toward my ultimate goal, which is to understand how human cells repair DNA.
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