title: 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 reannealing 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 labeling or surface immobilization. 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 nonspecificity of SSB binding to ssDNA further suggests that this approach could be used for fragment sizing of short ssDNA.
authors.affiliation: Department of Chemistry, Imperial College London , Exhibition Road, South Kensington Campus, London SW7 2AZ, U.K.
authors.forename: Deanpen
authors.initials: D
authors.lastname: Japrung
authors.forename: Azadeh
authors.initials: A
authors.lastname: Bahrami
authors.forename: Achim
authors.initials: A
authors.lastname: Nadzeyka
authors.forename: Lloyd
authors.initials: L
authors.lastname: Peto
authors.forename: Sven
authors.initials: S
authors.lastname: Bauerdick
authors.forename: Joshua B
authors.initials: JB
authors.lastname: Edel
authors.forename: Tim
authors.initials: T
authors.lastname: Albrecht
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D058608:Nanopores / Q000648:ultrastructure; D011485:Protein Binding
chemical_list: D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C476430:SSB protein, E coli