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. Author manuscript; available in PMC: 2022 Nov 1.
Published in final edited form as: Nat Struct Mol Biol. 2022 Apr 28;29(5):463–471. doi: 10.1038/s41594-022-00760-4

Fig. 1 |. H1 coalesces around nascent ssDNA.

Fig. 1 |

a, Schematic of the combined single-molecule fluorescence and force microscopy. A biotinylated λ-DNA molecule (48.5 kbp) is tethered between two streptavidin-coated polystyrene beads. b, A representative kymograph of Cy3-H1 binding to DNA over time as the inter-bead distance was increased. c, Total H1 signal across the DNA as a function of time for the kymograph shown in b. d, Distribution of the H1 signal along the DNA at two specific time points (T1 and T2) as indicated by the arrows in b. e, Cartoon illustrating the distinct binding configurations of H1 on DNA under different tensions. ssDNA is created by force-induced unpeeling. f, Schematic of two-color imaging for simultaneous visualization of H1 and RPA binding to DNA. g, A representative kymograph of Cy3-H1 (green) and AlexaFluor488-RPA (blue) binding to DNA over time as the inter-bead distance was increased. h, Total H1 and RPA signals across the DNA as a function of time for the kymograph shown in g. i, Distribution of the H1 (green) and RPA (blue) signals along the DNA at a specific time point (T1) as indicated by the arrow in g. j, Cartoon illustrating that H1 and RPA occupy separate regions of the tethered DNA. H1 coalesces around relaxed ssDNA, whereas RPA binds to ssDNA under tension.