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Abstract
Shelterin protein TRF2 modulates telomere structures by promoting dsDNA compaction and T-loop formation. Advancement of our understanding of the mechanism underlying TRF2-mediated DNA compaction requires additional information regarding DNA paths in TRF2-DNA complexes. To uncover the location of DNA inside protein-DNA complexes, we recently developed the Dual-Resonance-frequency-Enhanced Electrostatic force Microscopy (DREEM) imaging technique. DREEM imaging shows that in contrast to chromatin with DNA wrapping around histones, large TRF2-DNA complexes (with volumes larger than TRF2 tetramers) compact DNA inside TRF2 with portions of folded DNA appearing at the edge of these complexes. Supporting coarse-grained molecular dynamics simulations uncover the structural requirement and sequential steps during TRF2-mediated DNA compaction and result in folded DNA structures with protruding DNA loops as seen in DREEM imaging. Revealing DNA paths in TRF2 complexes provides new mechanistic insights into structure-function relationships underlying telomere maintenance pathways.
Highlights
Telomeres are nucleoprotein structures that prevent the degradation or fusion of the ends of linear chromosomes[1,2,3,4]
We recently developed an advanced imaging technique, Dual-Resonance-frequency-Enhanced Electrostatic force Microscopy (DREEM), which permits high-resolution imaging of weak electrostatic signals[32]
DREEM imaging using a DNA substrate containing both non-telomeric and telomeric regions (270 TTAGGG repeats) demonstrates that duplex DNA molecules are folded inside large TRF2 protein complexes with portions of the DNA appearing at the edge of the complexes
Summary
Telomeres are nucleoprotein structures that prevent the degradation or fusion of the ends of linear chromosomes[1,2,3,4]. DREEM imaging using a DNA substrate containing both non-telomeric and telomeric regions (270 TTAGGG repeats) demonstrates that duplex DNA molecules are folded inside large TRF2 protein complexes (with volume larger than tetramers) with portions of the DNA appearing at the edge of the complexes.
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