Abstract
Ultrafine anaphase bridges (UFBs) are thin DNA threads linking the separating sister chromatids in the anaphase of mitosis. UFBs are thought to form when topological DNA entanglements between two chromatids are not resolved prior to anaphase onset. In contrast to other markers of defective chromosome segregation, UFBs cannot be detected by direct staining of the DNA, but instead can be detected using immunofluorescence-based approaches. Due to the fact that they are short-lived and fragile in nature, UFBs can be challenging to detect. In this chapter, we describe methods that have been optimized for successful detection of UFBs. We also provide guidelines for the optimization of UFBs detection depending on the antibody and the cell line to be used.
The original version of this chapter was revised. An erratum to this chapter can be found at DOI 10.1007/978-1-4939-7306-4_43
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References
Chan KL, North PS, Hickson ID (2007) BLM is required for faithful chromosome segregation and its localization defines a class of ultrafine anaphase bridges. EMBO J 26(14):3397–3409. doi:10.1038/sj.emboj.7601777
Baumann C, Korner R, Hofmann K, Nigg EA (2007) PICH, a centromere-associated SNF2 family ATPase, is regulated by Plk1 and required for the spindle checkpoint. Cell 128(1):101–114. doi:10.1016/j.cell.2006.11.041
Wang LH, Schwarzbraun T, Speicher MR, Nigg EA (2008) Persistence of DNA threads in human anaphase cells suggests late completion of sister chromatid decatenation. Chromosoma 117(2):123–135. doi:10.1007/s00412-007-0131-7
Hengeveld RC, de Boer HR, Schoonen PM, de Vries EG, Lens SM, van Vugt MA (2015) Rif1 is required for resolution of ultrafine DNA bridges in anaphase to ensure genomic stability. Dev Cell 34(4):466–474. doi:10.1016/j.devcel.2015.06.014
Nielsen CF, Huttner D, Bizard AH, Hirano S, Li TN, Palmai-Pallag T, Bjerregaard VA, Liu Y, Nigg EA, Wang LH, Hickson ID (2015) PICH promotes sister chromatid disjunction and co-operates with topoisomerase II in mitosis. Nat Commun 6:8962. doi:10.1038/ncomms9962
Chan KL, Palmai-Pallag T, Ying S, Hickson ID (2009) Replication stress induces sister-chromatid bridging at fragile site loci in mitosis. Nat Cell Biol 11(6):753–760. doi:10.1038/ncb1882
Nielsen CF, Hickson ID (2016) PICH promotes mitotic chromsome segregation: identification of a novel role in ribosomal DNA disjunction. Cell Cycle 15(20):2704–2711
Barefield C, Karlseder J (2012) The BLM helicase contributes to telomere maintenance through processing of late-replicating intermediate structures. Nucleic Acids Res 40(15):7358–7367. doi:10.1093/nar/gks407
Vinciguerra P, Godinho SA, Parmar K, Pellman D, D’Andrea AD (2010) Cytokinesis failure occurs in Fanconi anemia pathway-deficient murine and human bone marrow hematopoietic cells. J Clin Invest 120(11):3834–3842. doi:10.1172/JCI43391
Nera B, Huang HS, Lai T, Xu L (2015) Elevated levels of TRF2 induce telomeric ultrafine anaphase bridges and rapid telomere deletions. Nat Commun 6:10132. doi:10.1038/ncomms10132
Kaulich M, Cubizolles F, Nigg EA (2012) On the regulation, function, and localization of the DNA-dependent ATPase PICH. Chromosoma 121(4):395–408. doi:10.1007/s00412-012-0370-0
Yankiwski V, Marciniak RA, Guarente L, Neff NF (2000) Nuclear structure in normal and Bloom syndrome cells. Proc Natl Acad Sci U S A 97(10):5214–5219. doi:10.1073/pnas.090525897
Acknowledgments
Work in the authors’ laboratory is supported by The European Research Council and The Danish National Research Foundation (grant DNRF115).
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Bizard, A.H., Nielsen, C.F., Hickson, I.D. (2018). Detection of Ultrafine Anaphase Bridges. In: Muzi-Falconi, M., Brown, G. (eds) Genome Instability. Methods in Molecular Biology, vol 1672. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7306-4_33
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DOI: https://doi.org/10.1007/978-1-4939-7306-4_33
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