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. 2020 May 13;117(21):11257–11264. doi: 10.1073/pnas.1920368117

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Fig. 6. — Nucleation of ScDmc1 nucleoprotein filament assembly is stimulated by ScRad51, but not by EcRecA. In the presence of short discontinuous patches of ScRad51 on ssDNA, assembly of ScDmc1 is stimulated. (A) In 351/dT90 DNA substrate, exemplary BM time course of ScDmc1 assembly and the nucleation time histogram show the nucleation time of 82.4 s. (B) Experiments of preformed ScRad51 patches on 351/ss(40+41)/Rad51 DNA substrates show the reduced nucleation time of 49.7 s. (C) Experiments of preformed EcRecA patches on 351/ss(40+41)/RecA DNA substrates show the longer nucleation time of 110.0 s. (D–E) His₆-tagged ScDmc1 incubated with ScRad51 (D) or EcRecA (E) was pulled down by TALON resins followed by wash and SDS elution. The supernatant (S), wash (W), and elution (E) were resolved by 12% SDS/PAGE and stained with Coomassie blue. (F) Proposed model for assembly of Dmc1 during nucleoprotein filament formation. The error bars of nucleation time are the SD by bootstrapping 5,000 times.