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Updated: Jan 19, 2026

CD Spectroscopy to Study DNA-Protein Interactions
Published on: February 10, 2022
Molecular determinants of Smc5/6 association with DNA junctions
Jeremy T-H Chang1,2, Victoria Miller-Browne3,4, Gabriella N L Chua1,5
1Laboratory of Nanoscale Biophysics and Biochemistry, The Rockefeller University, New York, NY, USA.
None:
Smc5/6 is an essential genome maintenance complex that interacts with double-stranded (ds) DNA, single-stranded (ss) DNA, and ss-dsDNA junctions. DNA association underlies Smc5/6's functions in managing intermediates generated during genome replication and repair. However, the mechanisms of this activity are not fully understood. Here, we report a single-molecule study examining Smc5/6 association with a dsDNA substrate containing a ssDNA gap with defined 3' and 5' junctions. We found that Smc5/6 associates with both 3' and 5' junctions but prefers the 3' junction in the presence of the ssDNA-binding complex RPA. Further, Smc5/6's junction association frequency and dwell time are regulated by two non-SMC subcomplexes and DNA binding residues of Smc6. Moreover, Smc5/6 prefers binding to junction sites free of the sliding clamp PCNA over those occupied with it. These results suggest that Smc5/6 utilizes its multiple structural modules to associate with junction sites in coordination with other genome maintenance factors.
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