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Pot1 OB-fold mutations unleash telomere instability to initiate tumorigenesis

Oncogene volume 36, pages 1939–1951 (2017)Cite this article

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Abstract

Chromosomal aberrations are a hallmark of human cancers, with complex cytogenetic rearrangements leading to genetic changes permissive for cancer initiation and progression. Protection of Telomere 1 (POT1) is an essential component of the shelterin complex and functions to maintain chromosome stability by repressing the activation of aberrant DNA damage and repair responses at telomeres. Sporadic and familial mutations in the oligosaccharide-oligonucleotide (OB) folds of POT1 have been identified in many human cancers, but the mechanism underlying how hPOT1 mutations initiate tumorigenesis has remained unclear. Here we show that the human POT1’s OB-folds are essential for the protection of newly replicated telomeres. Oncogenic mutations in hPOT1 OB-fold fail to bind to single-stranded telomeric DNA, eliciting a DNA damage response at telomeres that promote inappropriate chromosome fusions via the mutagenic alternative non-homologous end joining (A-NHEJ) pathway. hPOT1 mutations also result in telomere elongation and the formation of transplantable hematopoietic malignancies. Strikingly, conditional deletion of both mPot1a and p53 in mouse mammary epithelium resulted in development of highly invasive breast carcinomas and the formation of whole chromosomes containing massive arrays of telomeric fusions indicative of multiple breakage-fusion-bridge cycles. Our results reveal that hPOT1 OB-folds are required to protect and prevent newly replicated telomeres from engaging in A-NHEJ mediated fusions that would otherwise promote genome instability to fuel tumorigenesis.

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Acknowledgements

We thank Dr Asha Multani (MDACC, Houston, TX) for SKY analyses and Dr James You (MDACC, Houston, TX) for histological analyses and Dr Christos Hatzis (Yale university School of Medicine) for POT1 mutation analyses. We thank the Chang lab for helpful suggestions. This work was supported by the NCI (RO1 CA129037, R01 CA202816, R21 CA200506 and R21 CA182280) and the CT Department of Public Health (15-002167) to Sandy Chang. Support for Jayakrishnan Nandakumar is from the NIH (NIH R00-CA-167644-03, NIH R01-AG050509 (Jayakrishnan Nandakumar, co-investigator).

Author contributions

Sandy Chang conceived the project. Sandy Chang, Peili Gu and Jayakrishnan Nandakumar designed the experiments. Peili Gu, Yang Wang performed all the telomere biology and transplantation experiments. Ling Wu and Yang Xiao generated the breast cancer mouse model and performed tumour analyses. Kamlesh K. Bisht and Eric M. Smith performed the in vitro telomerase assay and Susan Bailey analysed the cytogenetics data. Peili Gu, Jayakrishnan Nandakumar, Ming Lei and Sandy Chang analysed and interpreted the data and composed the figures. Sandy Chang and Peili Gu wrote the paper.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT, USA

    P Gu, Y Wang, L Kukova & S Chang

  2. Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA

    K K Bisht, E M Smith & J Nandakumar

  3. Program in Chemical Biology, University of Michigan, Ann Arbor, MI, USA

    K K Bisht, E M Smith & J Nandakumar

  4. Department of GI Medical Oncology, UT MD Anderson Cancer Center, Houston, TX, USA

    L Wu

  5. Department of Medicine and Molecular and Cellular Biology, Section of Hematology-Oncology, Baylor College of Medicine, Houston, TX, USA

    Y Xiao

  6. Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA

    S M Bailey

  7. National Center for Protein Science Shanghai, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    M Lei

  8. Department of Pathology, Yale University School of Medicine, New Haven, CT, USA

    S Chang

  9. Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT, USA

    S Chang

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  1. P Gu
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Correspondence to S Chang.

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Gu, P., Wang, Y., Bisht, K. et al. Pot1 OB-fold mutations unleash telomere instability to initiate tumorigenesis. Oncogene 36, 1939–1951 (2017). https://doi.org/10.1038/onc.2016.405

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