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• W2997297854 abstract "Abstract RAG endonuclease initiates V(D)J recombination in progenitor (pro)-B cells 1 . Upon binding a recombination center (RC)-based J H , RAG scans upstream chromatin via loop extrusion, potentially mediated by cohesin 2–10 , to locate Ds and assemble a DJ H -based RC 11 . CTCF looping factor-bound elements (CBEs) within IGCR1 upstream of Ds impede RAG-scanning 12–15 ; but their inactivation allows scanning to proximal V H s where additional CBEs activate rearrangement and impede scanning any further upstream 15, 16 . Distal V H utilization is thought to involve diffusional RC access following large-scale Igh locus contraction 17–23 . Here, we test the potential of linear RAG-scanning to mediate distal V H usage in G1-arrested, v-Abl- pro-B cell lines 24, 25 , which undergo robust D-to-J H but little V H -to-DJ H rearrangements, presumably due to lack of locus contraction 11, 15 . Through an auxin-inducible approach 26, 27 , we degrade the cohesin-component Rad21 4, 7, 27 or CTCF 7, 9 in these G1-arrested lines, which maintain substantial viability throughout four-day experiments. Rad21 degradation eliminated all V(D)J recombination and RAG-scanning-associated interactions, except RC-located DQ52-to-J H joining in which synapsis occurs by diffusion 11 . Remarkably, while CTCF degradation suppressed most CBE-based chromatin interactions, it promoted robust RC interactions with, and robust V H -to-DJ H joining of, distal V H s, with patterns similar to those of “locus-contracted” primary pro-B cells. Thus, down-modulation of CTCF-bound scanning-impediment activity promotes cohesin-driven RAG-scanning across the 2.7Mb Igh locus." @default.
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• W2997297854 date "2020-01-02" @default.
• W2997297854 modified "2023-09-30" @default.
• W2997297854 title "CTCF orchestrates long-range cohesin-driven V(D)J recombinational scanning" @default.
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• W2997297854 doi "https://doi.org/10.1101/2020.01.01.891473" @default.
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