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<p>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.</p>

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Molecular determinants of Smc5/6 association with DNA junctions

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年1月17日

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  • Jeremy T-H. Chang,
  • Victoria Miller-Browne,
  • Gabriella N. L. Chua,
  • Jian Zheng,
  • Emily C. Beckwitt,
  • Shibai Li,
  • Bryce J. Katch,
  • Michael E. O’Donnell,
  • Shixin Liu,
  • Xiaolan Zhao

摘要

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.