Dynamic CTCF bridges drive genome organization
Caldwell, C. C.; Sa, M.; Hoogland, B. J.; Ridolfi, A.; Zaghal, N. F.; Broedersz, C. P.; Biebricher, A. S.; Wuite, G. J. L.
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Human CCCTC-binding factor (CTCF) is a crucial factor in genome organization, regulating chromatin looping and gene expression. The loop extrusion model (LEM) designates cohesin as the sole active player and limits CTCF to a passive barrier, though emerging evidence suggests a more dynamic role. Using complementary single-molecule techniques, combining dual- and quad-trap optical tweezers, fluorescence microscopy, and atomic force microscopy, we show CTCF forms stable, dynamic DNA bridges. We demonstrate CTCF binds with high affinity and cooperativity, undergoes 1D diffusion, and stiffens DNA. Strikingly, CTCF stabilizes DNA loops without cohesin and forms bridges that slide at low force yet resist rupture at forces exceeding dsDNA stability. These findings suggest a modified LEM in which CTCF directly forms dynamic, force-resistant bridges to stabilize cohesin-established loops, revealing an active role in genome organization.
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