Direct observation and analysis of the dynamics of the photoresponsive transcription factor GAL4

Abstract

Herein, the direct visualization of the dynamic interaction between a photoresponsive transcription factor fusion, GAL4–VVD, and DNA using high‐speed atomic force microscopy (HS‐AFM) is reported. A series of different GAL4–VVD movements, such as binding, sliding, stalling, and dissociation, was observed. Inter‐strand jumping on two double‐stranded (ds) DNAs was also observed. Detailed analysis using a long substrate DNA strand containing five GAL4‐binding sites revealed that GAL4–VVD randomly moved on the dsDNA using sliding and hopping to rapidly find specific binding sites, and then stalled to the specific sites to form a stable complex formation. These results suggest the existence of different conformations of the protein to enable sliding and stalling. This single‐molecule imaging system with nanoscale resolution provides an insight into the searching mechanism used by DNA‐binding proteins.