Multiple hPOT1–TPP1 cooperatively unfold contiguous telomeric G-quadruplexes proceeding from 3′ toward 5′, a feature due to a 3′-end binding preference and to structuring of telomeric DNA

Jean Chatain (1) , Georges Hatem (1) , Emmanuelle Delagoutte (1) , Jean-François Riou (1) , Patrizia Alberti (1) , Carole Saintomé (1)

1 STRING - Structure et Instabilité des Génomes

Telomeres are DNA repeated sequences that associate with shelterin proteins and protect the ends of eukaryotic chromosomes. Human telomeres are composed of 5 TTAGGG repeats and ends with a 3 single-stranded tail, called G-overhang, that can be specifically bound by the shelterin protein hPOT1 (human Protection of Telomeres 1). In vitro studies have shown that the telomeric G-strand can fold into stable contiguous G-quadruplexes (G4). In the present study we investigated how hPOT1, in complex with its shelterin partner TPP1, binds to telomeric sequences structured into contiguous G4 in potassium solutions. We observed that binding of multiple hPOT1-TPP1 preferentially proceeds from 3 toward 5. We explain this directionality in terms of two factors: (i) the preference of hPOT1-TPP1 for the binding site situated at the 3 end of a telomeric sequence and (ii) the cooperative binding displayed by hPOT1-TPP1 in potassium. By comparing binding in K + and in Li + , we demonstrate that this cooperative behaviour does not stem from protein-protein interactions, but from structuring of the telomeric DNA substrate into contiguous G4 in potassium. Our study suggests that POT1-TPP1, in physiological conditions, might preferentially cover the telomeric G-overhang starting from the 3-end and proceeding toward 5 .

Domains: Biochemistry [q-bio.BM]; Molecular biology; Biophysics

Jean Chatain, Georges Hatem, Emmanuelle Delagoutte, Jean-François Riou, Patrizia Alberti, et al.. Multiple hPOT1–TPP1 cooperatively unfold contiguous telomeric G-quadruplexes proceeding from 3′ toward 5′, a feature due to a 3′-end binding preference and to structuring of telomeric DNA. Nucleic Acids Research, 2021, 49, pp.10735 - 10746. ⟨10.1093/nar/gkab768⟩. ⟨mnhn-03383266⟩