@article{f3e82b119a4f4fae87b12e0d83943798,
title = "Compartmentalization of telomeres through DNA-scaffolded phase separation",
abstract = "Telomeres form unique nuclear compartments that prevent degradation and fusion of chromosome ends by recruiting shelterin proteins and regulating access of DNA damage repair factors. To understand how these dynamic components protect chromosome ends, we combine in vivo biophysical interrogation and in vitro reconstitution of human shelterin. We show that shelterin components form multicomponent liquid condensates with selective biomolecular partitioning on telomeric DNA. Tethering and anomalous diffusion prevent multiple telomeres from coalescing into a single condensate in mammalian cells. However, telomeres coalesce when brought into contact via an optogenetic approach. TRF1 and TRF2 subunits of shelterin drive phase separation, and their N-terminal domains specify interactions with telomeric DNA in vitro. Telomeric condensates selectively recruit telomere-associated factors and regulate access of DNA damage repair factors. We propose that shelterin mediates phase separation of telomeric chromatin, which underlies the dynamic yet persistent nature of the end-protection mechanism.",
keywords = "DNA repair, chromatin organization, phase separation, shelterin, telomeres",
author = "Amanda Jack and Yoonji Kim and Strom, {Amy R.} and Lee, {Daniel S.W.} and Byron Williams and Schaub, {Jeffrey M.} and Kellogg, {Elizabeth H.} and Finkelstein, {Ilya J.} and Ferro, {Luke S.} and Ahmet Yildiz and Brangwynne, {Clifford P.}",
note = "Funding Information: A.J., Y.K., and D.S.W.L. are supported by the NSF GRFP fellowship ( DGE-1752814 , A.J.; DGE-2039656 , Y.K.; DCE-1656466 , D.S.W.L.). Y.K. was previously supported by the NIGMS ( T32GM007388 ) while conducting experiments. A.R.S. is a Life Science Research Fellow through the Mark Foundation for Cancer Research ( AWD1006303 ). L.S.F. was supported by the NIH F32 Fellowship ( GM123655 ). This work was supported by NSF ( MCB-1617028 , A.Y.), NIGMS ( GM 118773 , A.Y.; GM120554 , I.J.F., and GM124463 , E.H.K.), the NIH 4D Nucleome Program ( U01 DA040601 , C.P.B.), CPRIT ( RP190301 to I.J.F.), the Welch Foundation ( F-1808 to I.J.F.), and the Howard Hughes Medical Institute (C.P.B.). The content is solely the responsibility of the authors and does not represent the official views of these funding institutions. Funding Information: We thank Joshua Riback, Jorine Eeftens, Yi-Che Chang, David Sanders, Evangelos Gatzogiannis, Yavuz S. Dagdas, John T. Canty, and other members of the Yildiz and Brangwynne laboratories for helpful discussions, Shunsuke Shimobayashi for the FUSN-miRFP-TRF1 construct, David Sanders for the FM5 vectors, Yi-Che Chang for helpful discussions on data analysis, Sofi Quinodoz for help with the control siRNA transfection efficiency experiment, Lindsay Becker for help with the immunofluorescence protocol, Titia de Lange (Rockefeller Univ.) for the hTERT-RPE1 cell line, Huaiying Zhang (Carnegie Mellon Univ.) for the HeLa RMCE GFP-TRF1 cell line, UC Berkeley MacroLab for the TEV protease and competent cells, and UC Berkeley Cell Culture Facility for the insect cell cultures. A.J. Y.K. and D.S.W.L. are supported by the NSF GRFP fellowship (DGE-1752814, A.J.; DGE-2039656, Y.K.; DCE-1656466, D.S.W.L.). Y.K. was previously supported by the NIGMS (T32GM007388) while conducting experiments. A.R.S. is a Life Science Research Fellow through the Mark Foundation for Cancer Research (AWD1006303). L.S.F. was supported by the NIH F32 Fellowship (GM123655). This work was supported by NSF (MCB-1617028, A.Y.), NIGMS (GM 118773, A.Y.; GM120554, I.J.F. and GM124463, E.H.K.), the NIH 4D Nucleome Program (U01 DA040601, C.P.B.), CPRIT (RP190301 to I.J.F.), the Welch Foundation (F-1808 to I.J.F.), and the Howard Hughes Medical Institute (C.P.B.). The content is solely the responsibility of the authors and does not represent the official views of these funding institutions. A.J. Y.K. A.R.S. L.S.F. D.S.W.L. A.Y. and C.P.B. conceived of experiments. A.J. and L.S.F. cloned shelterin constructs and purified the protein. A.J. performed the in vitro experiments and analyzed the data. L.K. and B.W. purified nucleosomes, and J.M.S. and I.J.F. purified RPA. A.J. Y.K. and A.R.S. cloned constructs, and Y.K. curated cell lines. Y.K. and A.R.S. performed live-cell imaging. Y.K. A.R.S. and D.S.W.L. analyzed live-cell imaging data. A.J. Y.K. A.R.S. D.S.W.L. L.S.F. A.Y. and C.P.B. wrote the manuscript with input from all authors. C.P.B. is a founder of and consultant for Nereid Therapeutics. Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
month = jan,
day = "24",
doi = "10.1016/j.devcel.2021.12.017",
language = "English (US)",
volume = "57",
pages = "277--290.e9",
journal = "Developmental Cell",
issn = "1534-5807",
publisher = "Cell Press",
number = "2",
}