{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T21:07:03Z","timestamp":1761599223649,"version":"3.37.3"},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"18","license":[{"start":{"date-parts":[[2021,9,17]],"date-time":"2021-09-17T00:00:00Z","timestamp":1631836800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007522","name":"Mus\u00e9um National d\u2019Histoire Naturelle","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100007522","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004794","name":"Centre National de la Recherche Scientifique","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004794","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001677","name":"Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001677","id-type":"DOI","asserted-by":"publisher"}]},{"name":"MNHN Graduate School","award":["ED 227"],"award-info":[{"award-number":["ED 227"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,10,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Telomeres are DNA repeated sequences that associate with shelterin proteins and protect the ends of eukaryotic chromosomes. Human telomeres are composed of 5\u2032TTAGGG repeats and ends with a 3\u2032 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\u2013TPP1 preferentially proceeds from 3\u2032 toward 5\u2032. We explain this directionality in terms of two factors: (i) the preference of hPOT1\u2013TPP1 for the binding site situated at the 3\u2032 end of a telomeric sequence and (ii) the cooperative binding displayed by hPOT1\u2013TPP1 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\u2032-end and proceeding toward 5\u2032.<\/jats:p>","DOI":"10.1093\/nar\/gkab768","type":"journal-article","created":{"date-parts":[[2021,9,15]],"date-time":"2021-09-15T11:17:00Z","timestamp":1631704620000},"page":"10735-10746","source":"Crossref","is-referenced-by-count":5,"title":["Multiple hPOT1\u2013TPP1 cooperatively unfold contiguous telomeric G-quadruplexes proceeding from 3\u2032 toward 5\u2032, a feature due to a 3\u2032-end binding preference and to structuring of telomeric DNA"],"prefix":"10.1093","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0703-9181","authenticated-orcid":false,"given":"Jean","family":"Chatain","sequence":"first","affiliation":[{"name":"Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um national d\u2019Histoire naturelle, CNRS, INSERM, 43 rue Cuvier, F-75005\u00a0Paris, France"}]},{"given":"Georges","family":"Hatem","sequence":"additional","affiliation":[{"name":"Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um national d\u2019Histoire naturelle, CNRS, INSERM, 43 rue Cuvier, F-75005\u00a0Paris, France"}]},{"given":"Emmanuelle","family":"Delagoutte","sequence":"additional","affiliation":[{"name":"Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um national d\u2019Histoire naturelle, CNRS, INSERM, 43 rue Cuvier, F-75005\u00a0Paris, France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0055-6506","authenticated-orcid":false,"given":"Jean-Fran\u00e7ois","family":"Riou","sequence":"additional","affiliation":[{"name":"Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um national d\u2019Histoire naturelle, CNRS, INSERM, 43 rue Cuvier, F-75005\u00a0Paris, France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5231-3768","authenticated-orcid":false,"given":"Patrizia","family":"Alberti","sequence":"additional","affiliation":[{"name":"Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um national d\u2019Histoire naturelle, CNRS, INSERM, 43 rue Cuvier, F-75005\u00a0Paris, France"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7609-6913","authenticated-orcid":false,"given":"Carole","family":"Saintom\u00e9","sequence":"additional","affiliation":[{"name":"Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um national d\u2019Histoire naturelle, CNRS, INSERM, 43 rue Cuvier, F-75005\u00a0Paris, France"},{"name":"Sorbonne Universit\u00e9, UFR927, F-75005\u00a0Paris, France"}]}],"member":"286","published-online":{"date-parts":[[2021,9,17]]},"reference":[{"key":"2021100913443791600_B1","doi-asserted-by":"crossref","first-page":"2100","DOI":"10.1101\/gad.1346005","article-title":"Shelterin: The protein complex that shapes and safeguards human telomeres","volume":"19","author":"de\u00a0Lange","year":"2005","journal-title":"Genes Dev."},{"key":"2021100913443791600_B2","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1016\/j.febslet.2004.11.036","article-title":"Telomeres and telomerase: their mechanisms of action and the effects of altering their functions","volume":"579","author":"Blackburn","year":"2005","journal-title":"FEBS Lett."},{"key":"2021100913443791600_B3","first-page":"703","article-title":"G-quartet structures in telomeric DNA","volume":"23","author":"Williamson","year":"1994","journal-title":"Annu. Rev."},{"key":"2021100913443791600_B4","doi-asserted-by":"crossref","first-page":"3686","DOI":"10.3390\/molecules25163686","article-title":"G-quadruplexes at telomeres: friend or foe","volume":"25","author":"Bryan","year":"2020","journal-title":"Molecules"},{"key":"2021100913443791600_B5","doi-asserted-by":"crossref","first-page":"10864","DOI":"10.1073\/pnas.0504744102","article-title":"Human POT1 disrupts telomeric G-quadruplexes allowing telomerase extension in vitro","volume":"102","author":"Zaug","year":"2005","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021100913443791600_B6","doi-asserted-by":"crossref","first-page":"17347","DOI":"10.1073\/pnas.0707365104","article-title":"Reevaluation of telomerase inhibition by quadruplex ligands and their mechanisms of action","volume":"104","author":"De\u00a0Cian","year":"2007","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021100913443791600_B7","doi-asserted-by":"crossref","first-page":"6229","DOI":"10.1093\/nar\/gkr164","article-title":"G-quadruplex formation at the 3\u2032 end of telomere DNA inhibits its extension by telomerase, polymerase and unwinding by helicase","volume":"39","author":"Wang","year":"2011","journal-title":"Nucleic Acids Res."},{"key":"2021100913443791600_B8","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1016\/j.str.2014.03.013","article-title":"Telomeric overhang length determines structural dynamics and accessibility to telomerase and ALT-associated proteins","volume":"22","author":"Hwang","year":"2014","journal-title":"Structure"},{"key":"2021100913443791600_B9","doi-asserted-by":"crossref","first-page":"11752","DOI":"10.1093\/nar\/gkx789","article-title":"Molecular mechanisms by which oxidative DNA damage promotes telomerase activity","volume":"45","author":"Lee","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"2021100913443791600_B10","doi-asserted-by":"crossref","first-page":"1171","DOI":"10.1126\/science.1060036","article-title":"Pot1, the putative telomere end-binding protein in fission yeast and humans","volume":"292","author":"Baumann","year":"2001","journal-title":"Science"},{"key":"2021100913443791600_B11","doi-asserted-by":"crossref","first-page":"8079","DOI":"10.1128\/MCB.22.22.8079-8087.2002","article-title":"Human pot1 (protection of telomeres) protein: Cytolocalization, gene structure, and alternative splicing","volume":"22","author":"Baumann","year":"2002","journal-title":"Mol. Cell. Biol."},{"key":"2021100913443791600_B12","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1038\/nature01688","article-title":"POT1 as a terminal transducer of TRF1 telomere length control","volume":"423","author":"Loayza","year":"2003","journal-title":"Nature"},{"key":"2021100913443791600_B13","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1016\/S0960-9822(03)00339-7","article-title":"Human POT1 facilitates telomere elongation by telomerase","volume":"13","author":"Colgin","year":"2003","journal-title":"Curr. Biol."},{"key":"2021100913443791600_B14","doi-asserted-by":"crossref","first-page":"3552","DOI":"10.1128\/MCB.24.8.3552-3561.2004","article-title":"Rescue of an hTERT mutant defective in telomere elongation by fusion with hPot1","volume":"24","author":"Armbruster","year":"2004","journal-title":"Mol. Cell. Biol."},{"key":"2021100913443791600_B15","doi-asserted-by":"crossref","first-page":"20449","DOI":"10.1074\/jbc.M502212200","article-title":"Switching human telomerase on and off with hPOT1 protein in vitro","volume":"280","author":"Lei","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2021100913443791600_B16","doi-asserted-by":"crossref","first-page":"1223","DOI":"10.1038\/nsmb867","article-title":"Structure of human POT1 bound to telomeric single-stranded DNA provides a model for chromosome end-protection","volume":"11","author":"Lei","year":"2004","journal-title":"Nat. Struct. Mol. Biol."},{"key":"2021100913443791600_B17","doi-asserted-by":"crossref","first-page":"13241","DOI":"10.1074\/jbc.M312309200","article-title":"DNA binding features of human POT1","volume":"279","author":"Loayza","year":"2004","journal-title":"J. Biol. Chem."},{"key":"2021100913443791600_B18","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1038\/ncb1142","article-title":"PTOP interacts with POT1 and regulates its localization to telomeres","volume":"6","author":"Liu","year":"2004","journal-title":"Nat. Cell Biol."},{"key":"2021100913443791600_B19","doi-asserted-by":"crossref","first-page":"1649","DOI":"10.1101\/gad.1215404","article-title":"POT1-interacting protein PIP1: a telomere length regulator that recruits POT1 to the TIN2\/TRF1 complex","volume":"18","author":"Ye","year":"2004","journal-title":"Genes Dev."},{"key":"2021100913443791600_B20","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1038\/nature05454","article-title":"The POT1-TPP1 telomere complex is a telomerase processivity factor","volume":"445","author":"Wang","year":"2007","journal-title":"Nature"},{"key":"2021100913443791600_B21","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1038\/nature05469","article-title":"TPP1 is a homologue of ciliate TEBP-beta and interacts with POT1 to recruit telomerase","volume":"445","author":"Xin","year":"2007","journal-title":"Nature"},{"key":"2021100913443791600_B22","doi-asserted-by":"crossref","first-page":"1872","DOI":"10.1016\/j.str.2012.08.018","article-title":"POT1-TPP1 regulates telomeric overhang structural dynamics","volume":"20","author":"Hwang","year":"2012","journal-title":"Structure"},{"key":"2021100913443791600_B23","doi-asserted-by":"crossref","first-page":"2990","DOI":"10.1073\/pnas.1321436111","article-title":"G-quadruplex formation in telomeres enhances POT1\/TPP1 protection against RPA binding","volume":"111","author":"Ray","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2021100913443791600_B24","doi-asserted-by":"crossref","first-page":"2695","DOI":"10.1016\/j.jmb.2016.04.031","article-title":"POT1-TPP1 binding and unfolding of telomere DNA discriminates against structural polymorphism","volume":"428","author":"Mullins","year":"2016","journal-title":"J. Mol. Biol."},{"key":"2021100913443791600_B25","doi-asserted-by":"crossref","first-page":"4976","DOI":"10.1093\/nar\/gkaa202","article-title":"Human POT1 unfolds G-quadruplexes by conformational selection","volume":"48","author":"Chaires","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"2021100913443791600_B26","doi-asserted-by":"crossref","first-page":"15461","DOI":"10.1021\/ja064536h","article-title":"Characterization of structure and stability of long telomeric DNA G-quadruplexes","volume":"128","author":"Yu","year":"2006","journal-title":"J. Am. Chem. Soc."},{"key":"2021100913443791600_B27","doi-asserted-by":"crossref","first-page":"2871","DOI":"10.1021\/ja807313x","article-title":"Human telomeric DNA sequence-specific cleaving by G-quadruplex formation","volume":"131","author":"Xu","year":"2009","journal-title":"J. Am. Chem. Soc."},{"key":"2021100913443791600_B28","doi-asserted-by":"crossref","first-page":"20951","DOI":"10.1021\/ja209192a","article-title":"Structure and stability of higher-order human telomeric quadruplexes","volume":"133","author":"Petraccone","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"2021100913443791600_B29","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.biochi.2015.04.003","article-title":"Understanding the stability of DNA G-quadruplex units in long human telomeric strands","volume":"113","author":"Bugaut","year":"2015","journal-title":"Biochimie"},{"key":"2021100913443791600_B30","doi-asserted-by":"crossref","first-page":"7479","DOI":"10.1074\/jbc.M110.205641","article-title":"Single molecule studies of physiologically relevant telomeric tails reveal POT1 mechanism for promoting G-quadruplex unfolding","volume":"286","author":"Wang","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2021100913443791600_B31","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.jmb.2011.04.049","article-title":"MultiplePOT1-TPP1 proteins coat and compact long telomeric single-stranded DNA","volume":"410","author":"Taylor","year":"2011","journal-title":"J. Mol. Biol."},{"key":"2021100913443791600_B32","doi-asserted-by":"crossref","first-page":"16361","DOI":"10.1074\/jbc.M113.471896","article-title":"Coordinated interactions of multiple POT1-TPP1 proteins with telomere DNA","volume":"288","author":"Corriveau","year":"2013","journal-title":"J. Biol. Chem."},{"key":"2021100913443791600_B33","doi-asserted-by":"crossref","first-page":"23527","DOI":"10.1073\/pnas.1905381116","article-title":"POT1-TPP1 differentially regulates telomerase via POT1 his266 and as a function of single-stranded telomere DNA length","volume":"116","author":"Xu","year":"2019","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021100913443791600_B34","doi-asserted-by":"crossref","first-page":"1942","DOI":"10.15252\/embj.201490773","article-title":"RPA prevents g-rich structure formation at lagging-strand telomeres to allow maintenance of chromosome ends","volume":"34","author":"Audry","year":"2015","journal-title":"EMBO J."},{"key":"2021100913443791600_B35","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.biochi.2017.11.017","article-title":"The binding efficiency of RPA to telomeric g-strands folded into contiguous g-quadruplexes is independent of the number of G4 units","volume":"146","author":"Lancrey","year":"2018","journal-title":"Biochimie"},{"key":"2021100913443791600_B36","doi-asserted-by":"crossref","first-page":"17.1.1","DOI":"10.1002\/0471142700.nc1701s37","article-title":"UV Melting of G-Quadruplexes","volume":"37","author":"Mergny","year":"2009","journal-title":"Curr. Protoc. Nucleic Acid Chem."},{"key":"2021100913443791600_B37","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.biochi.2014.04.006","article-title":"Binding polarity of RPA to telomeric sequences and infuence of G-quadruplex stability","volume":"103","author":"Safa","year":"2014","journal-title":"Biochimie"},{"key":"2021100913443791600_B38","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1111\/j.1742-4658.2009.07464.x","article-title":"Human telomeric G-quadruplex: structures of DNA and RNA sequences","volume":"277","author":"Anh Tu\u00e2n Phan","year":"2010","journal-title":"FEBS Lett."},{"key":"2021100913443791600_B39","doi-asserted-by":"crossref","first-page":"2816","DOI":"10.1021\/ja312403b","article-title":"High-resolution insight into G-overhang architecture","volume":"135","author":"H\u00e4nsel","year":"2013","journal-title":"J. Am. Chem. Soc."},{"key":"2021100913443791600_B40","doi-asserted-by":"crossref","first-page":"2146","DOI":"10.1074\/jbc.M115.709667","article-title":"5\u2032 to 3\u2032 unfolding directionality of DNA secondary structures by replication protein A","volume":"291","author":"Safa","year":"2016","journal-title":"J. Biol. Chem."},{"key":"2021100913443791600_B41","doi-asserted-by":"crossref","first-page":"2058","DOI":"10.1021\/bi00006a028","article-title":"Recombinant human replication protein A binds to polynucleotides with low cooperativity","volume":"34","author":"Kim","year":"1995","journal-title":"Biochemistry"},{"key":"2021100913443791600_B42","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.molcel.2005.02.035","article-title":"Telomere-end processing the terminal nucleotides of human chromosomes","volume":"18","author":"Sfeir","year":"2005","journal-title":"Mol. Cell"}],"container-title":["Nucleic Acids Research"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/nar\/article-pdf\/49\/18\/10735\/40538261\/gkab768.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/nar\/article-pdf\/49\/18\/10735\/40538261\/gkab768.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,10,11]],"date-time":"2021-10-11T01:43:48Z","timestamp":1633916628000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/nar\/article\/49\/18\/10735\/6371971"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,17]]},"references-count":42,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2021,9,17]]},"published-print":{"date-parts":[[2021,10,11]]}},"URL":"https:\/\/doi.org\/10.1093\/nar\/gkab768","relation":{},"ISSN":["0305-1048","1362-4962"],"issn-type":[{"type":"print","value":"0305-1048"},{"type":"electronic","value":"1362-4962"}],"subject":[],"published-other":{"date-parts":[[2021,10,11]]},"published":{"date-parts":[[2021,9,17]]}}}