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A quantitative telomeric chromatin isolation protocol identifies different telomeric states

Author

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  • Larissa Grolimund

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    ISREC-Swiss Institute fro Experimental Cancer Research at EPFL)

  • Eric Aeby

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    ISREC-Swiss Institute fro Experimental Cancer Research at EPFL)

  • Romain Hamelin

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    Proteomics Core Facility at EPFL)

  • Florence Armand

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    Proteomics Core Facility at EPFL)

  • Diego Chiappe

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    Proteomics Core Facility at EPFL)

  • Marc Moniatte

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    Proteomics Core Facility at EPFL)

  • Joachim Lingner

    (EPFL-Ecole Polytechnique Fédérale de Lausanne, School of Life Sciences
    ISREC-Swiss Institute fro Experimental Cancer Research at EPFL)

Abstract

Telomere composition changes during tumourigenesis, aging and in telomere syndromes in a poorly defined manner. Here we develop a quantitative telomeric chromatin isolation protocol (QTIP) for human cells, in which chromatin is cross-linked, immunopurified and analysed by mass spectrometry. QTIP involves stable isotope labelling by amino acids in cell culture (SILAC) to compare and identify quantitative differences in telomere protein composition of cells from various states. With QTIP, we specifically enrich telomeric DNA and all shelterin components. We validate the method characterizing changes at dysfunctional telomeres, and identify and validate known, as well as novel telomere-associated polypeptides including all THO subunits, SMCHD1 and LRIF1. We apply QTIP to long and short telomeres and detect increased density of SMCHD1 and LRIF1 and increased association of the shelterins TRF1, TIN2, TPP1 and POT1 with long telomeres. Our results validate QTIP to study telomeric states during normal development and in disease.

Suggested Citation

  • Larissa Grolimund & Eric Aeby & Romain Hamelin & Florence Armand & Diego Chiappe & Marc Moniatte & Joachim Lingner, 2013. "A quantitative telomeric chromatin isolation protocol identifies different telomeric states," Nature Communications, Nature, vol. 4(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:4:y:2013:i:1:d:10.1038_ncomms3848
    DOI: 10.1038/ncomms3848
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    Cited by:

    1. Hanna Braun & Ziyan Xu & Fiona Chang & Nikenza Viceconte & Grishma Rane & Michal Levin & Liudmyla Lototska & Franziska Roth & Alexia Hillairet & Albert Fradera-Sola & Vartika Khanchandani & Zi Wayne S, 2023. "ZNF524 directly interacts with telomeric DNA and supports telomere integrity," Nature Communications, Nature, vol. 14(1), pages 1-17, December.

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