Single molecule analysis of DNA bending by the human mitochondrial transcription factor A (TFAM) (360G-Wellcome-207097_Z_17_Z)

Visualization of the dynamics of transcription initiation by mitochondrial RNA polymerase in real-time performed at single-molecule resolution forms the basis of this proposal. The human mitochondrial transcription factor A (TFAM) carries out DNA bending and my aim is to investigate whether this is an essential feature for transcription initiation by mitochondrial RNA polymerase. Multi-colour single-molecule super-resolution microscopy is the technique which will be used to demonstrate whether this DNA bending phenomenon is required for transcription initiation. This is a high-throughput technology capable of visualising reactions at single-molecule level, especially transcription factor binding, DNA bending and production of RNA. This state-of the-art instrument makes use of three different colours for accurate detection; green and red for smFRET and blue for RNA. I am also intrigued and have a desire to investigate for the first time whether DNA bending by TFAM is correlated with transcription by mtRNAP molecules.