title: Single molecule studies of physiologically relevant telomeric tails reveal POT1 mechanism for promoting G-quadruplex unfolding.
abstract: Human telomeres are composed of duplex TTAGGG repeats and a 3' single-stranded DNA tail. The telomeric DNA is protected and regulated by the shelterin proteins, including the protection of telomeres 1 (POT1) protein that binds telomeric single-stranded DNA. The single-stranded tail can fold into G-quadruplex (G4) DNA. Both POT1 and G4 DNA play important roles in regulating telomere length homeostasis. To date, most studies have focused on individual quadruplexes formed by four TTAGGG repeats. Telomeric tails in human cells have on average six times as many repeats, and no structural studies have examined POT1 binding in competition with G4 DNA folding. Using single molecule atomic force microscopy imaging, we observed that the majority of the telomeric tails of 16 repeats formed two quadruplexes even though four were possible. The result that physiological telomeric tails rarely form the maximum potential number of G4 units provides a structural basis for the coexistence of G4 and POT1 on the same DNA molecule, which is observed directly in the captured atomic force microscopy images. We further observed that POT1 is significantly more effective in disrupting quadruplex DNA on long telomeric tails than an antisense oligonucleotide, indicating a novel POT1 activity beyond simply preventing quadruplex folding.
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA. wangh4@upmc.edu
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Gerald J
authors.initials: GJ
authors.lastname: Nora
authors.forename: Harshad
authors.initials: H
authors.lastname: Ghodke
authors.forename: Patricia L
authors.initials: PL
authors.lastname: Opresko
mesh_terms: D000818:Animals; D002460:Cell Line; D020650:Combinatorial Chemistry Techniques; D004277:DNA, Single-Stranded / Q000737:chemistry*; D054856:G-Quadruplexes; D006801:Humans; D007313:Insecta; D018625:Microscopy, Atomic Force; D008956:Models, Chemical; D016376:Oligonucleotides, Antisense; D011485:Protein Binding; D000089804:Shelterin Complex; D016615:Telomere / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D034501:Telomere-Binding Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D004277:DNA, Single-Stranded; D016376:Oligonucleotides, Antisense; C440849:POT1 protein, human; D000089804:Shelterin Complex; D034501:Telomere-Binding Proteins