Real-Time Structural Biology of DNA and DNA-Protein Complexes on an Optical Microscope
Szalai, A. M.; Ferrari, G.; Richter, L.; Hartmann, J.; Kesici, M.-Z.; Ji, B.; Koshic, K.; Jaeger, A.; Aksimentiev, A.; Tessmer, I.; Kaminska, I.; Vera, A. M.; Tinnefeld, P.
Show abstract
The intricate interplay between DNA and proteins is key for biological functions such as DNA replication, transcription, and repair. To better understand these interactions, it is crucial to develop tools to study DNA-protein complexes with high spatial and temporal resolution. Here, we use the vertical orientation that DNA adopts on graphene and investigate its interactions with proteins via energy transfer from a probe dye to graphene, achieving spatial resolution down to the [A]ngstrom scale. We measured the bending angle of DNA induced by adenine tracts, bulges, abasic sites and the binding of Escherichia coli endonuclease IV with unprecedented precision and millisecond time resolution. Additionally, we observed the translocation of the O6-alkylguanine DNA alkyltransferase along double-stranded DNA, reaching single-base pair resolution and detecting an affinity for adenine tracts. Overall, we foresee that this method will become a widespread tool for the dynamical study of nucleic acid and nucleic acid-protein interactions.
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