SSB Binding to Single-Stranded DNA Probed Using Solid-State Nanopore Sensors

Abstract

Single stranded DNA (ssDNA) binding protein plays an important role in the DNA replication process in a wide range of organisms. It binds to ssDNA to prevent premature re annealing and to protect it from degradation. Current understanding of SSB/ssDNA interaction points to a complex mechanism, including SSB motion along the DNA strand. We report on the first characterization of this interaction at the single molecule level using solid state nanopore sensors, namely without any labelling or surface immobilisation. Our results show that the presence of SSB on the ssDNA can control the speed of nanopore translocation, presumably due to strong interactions between SSB and the nanopore surface. This enables nanopore based detection of ssDNA fragments as short as 37 nt, which is normally very difficult with solid state nanopore sensors, due to constraints in noise and bandwidth.Notably, this fragment is considerably shorter than the 65 nt binding motif, typically required for SSB binding at high salt concentrations. The non specificity of SSB binding to ssDNA further suggests that this approach could be used for fragment sizing of short ssDNA.

High-speed detection of DNA translocation in nanopipettes

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Self-Aligned Plasmonic Nanopores by Optically Controlled Dielectric Breakdown

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“…206, 207 Their studies demonstrated that the SSB– ssDNA complex has a characteristic current signature that is distinct from either SSB alone or ssDNA alone. Of note, SSB only binds to ssDNA but not dsDNA.…”