Direct visualization of the formation of RecA/dsDNA complexes at the single-molecule level.
The assembly of RecA on linear dsDNA with ATPγS in the reaction was elucidated using atomic force microscopy (AFM) on a single-molecule level. It was found that assembly generally (∼95%) proceeded from a single nucleation site that started from one end of the DNA strand. About 5% of the complexes were formed starting either from both ends or from the middle of dsDNA strand. In all these cases, the RecA coating was contiguous for each region suggesting the binding of RecA to DNA is cooperative. The AFM observation provides direct experimental evidence to show how RecA binds to linear dsDNA in the presence of ATPγS.
School of Chemistry and Chemical Engineering, Shenzhen University, University of Toronto, M5S 3H6 Toronto, Ontario, Canada. phbingsl@gmail.com
Bing Shi
BS
Li
Bo
B
Wei
M Cynthia
MC
Goh
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D011938:Rec A Recombinases / Q000737:chemistry* / Q000648:ultrastructure
C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D004247:DNA; D011938:Rec A Recombinases