Engineered holliday junctions as single-molecule reporters for protein-DNA interactions with application to a MerR-family regulator.

ABSTRACT: Protein-DNA interactions are essential for gene maintenance, replication, and expression. Characterizing how proteins interact with and change the structure of DNA is crucial in elucidating the mechanisms of protein function. Here, we present a novel and generalizable method of using engineered DNA Holliday junctions (HJs) that contain specific protein-recognition sequences to report protein-DNA interactions in single-molecule FRET measurements, utilizing the intrinsic structural dynamics of HJs. Because the effects of protein binding are converted to the changes in the structure and dynamics of HJs, protein-DNA interactions that involve small structural changes of DNA can be studied. We apply this method to investigate how the MerR-family regulator PbrR691 interacts with DNA for transcrip

SUBMITTER: Sarkar SK 

PROVIDER: S-EPMC2528078 | biostudies-literature | 2007 Oct

REPOSITORIES: biostudies-literature