Fig. 3: 5’-OH moieties and catalytically dead HaloTag-635-LIG3α (K421A), alter nick binding kinetics.
From: The zinc finger of DNA ligase 3α binds to nucleosomes via an arginine anchor
a A corresponding kymograph of an observation for HaloTag-635-LIG3α-XRCC1-YFP binding calf intestinal alkaline phosphatase (CIP) treated 10 nicked λ DNA generating 8 observable non-ligatable nick sites at 10 pN tension. b Cumulative residence time distribution (CRTD) and cumulative gap time distribution (CGTD), analysis for HaloTag-635-LIG3α binding CIP treated nicked λ DNA, with a single exponential fit shown in red. c Cumulative residence time distribution (CRTD) and cumulative gap time distribution (CGTD), analysis for XRCC1-YFP binding CIP treated nicked λ DNA, with a single exponential fit shown in blue. d The distribution of the 11 categories for HaloTag-635-LIG3α and XRCC1-YFP binding to CIP treated non-ligatable 10 nicked λ DNA. Error bars represent the SEM of two experiments. e 11 co-localization categories as key. f A representative structural model of catalytically dead N-terminal tagged HaloTag-635-K421A LIG3α bound to nicked DNA (PDB code 3L2P). g A corresponding kymograph of an observation for HaloTag-635-K421A LIG3α-XRCC1-YFP binding to10 nicked λ DNA at 10 pN tension. h Cumulative residence time distribution (CRTD) and cumulative gap time distribution (CGTD), analysis for HaloTag-635-K421A-LIG3α, with a single exponential fit shown in red. i Cumulative residence time distribution (CRTD) and cumulative gap time distribution (CGTD), analysis for XRCC1-YFP, with a single exponential fit shown in blue. j The distribution of the 11 categories for HaloTag-635- K421A LIG3α and XRCC1-YFP binding to ligatable 10 nicked λ DNA. Error bars represent the SEM of two experiments. Created in BioRender. Gupta, A. (2025) https://BioRender.com/ hcn8qmv. Source data are provided as a Source Data file.
