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PreprintCited by 0Microtubule and mitosis dynamicsDNA Repair MechanismsGenomics and Chromatin Dynamics

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Centromere protection requires strict mitotic inactivation of the Bloom syndrome helicase complex

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Chan, Kok-Lung, Fernández-Casañas, María, Karanika, Eleftheria, Olukoga, Tomisin, Herbert, Alex, Aliyaskarova, Umit, Day, Matthew·2024·Research Square
DOI 10.21203/rs.3.rs-4420374/v1

Preprint — published as 10.1038/s41467-025-62966-6 · Paper page

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Abstract

The BTRR (BLM/TOP3A/RMI1/RMI2) complex resolves various DNA replication and recombination intermediates to suppress genome instability. Alongside PICH, they target mitotic DNA intertwinements, known as ultrafine DNA bridges, facilitating chromosome segregation. Both BLM and PICH undergo transient mitotic hyper-phosphorylation, but the biological significance of this remains elusive. Here, we uncover that during early mitosis, multiple protein kinases act together to strictly constrain the BTRR complex for the protection of centromeres. Mechanistically, CDK1 destabilises the complex and suppresses its association with PICH at the chromatin underneath kinetochores. Inactivating the BLM and TOP3A interaction compromises the UFB-binding complex mitotic functions and can prevent centromere destruction. We further unravel how different clusters of mitotic phosphorylation on BLM affect its interaction with the TOP3A/RMI1/RMI2 subcomplex and illegitimate centromere unwinding. Furthermore, we identify specific phosphorylation sites targeted by the MPS1-PLK1 axis functioning to prevent BLM hyper-activation at centromeres. Notably, unleashing such activity after sister-chromatid cohesion loss facilitates separation of entangled chromosomes. Together, our study defines a centromere protection pathway in human mitotic cells, heavily reliant on a tight spatiotemporal control of the BTRR complex.

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  1. 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 26:3397–3409. https://doi.org/10.1038/sj.emboj.7601777 · DOI
  2. 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 128:101–114. https://doi.org/10.1016/j.cell.2006.11.041 · DOI
  3. 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 11:753–760. https://doi.org/10.1038/ncb1882 · DOI
  4. Chan YW, Fugger K, West SC (2018) Unresolved recombination intermediates lead to ultra-fine anaphase bridges, chromosome breaks and aberrations. Nat Cell Biol 20:92–103. https://doi.org/10.1038/s41556-017-0011-1 · DOI
  5. Tiwari A, Addis Jones O, Chan K (2018) L. 53BP1 can limit sister-chromatid rupture and rearrangements driven by a distinct ultrafine DNA bridging-breakage process. Nat Commun 9:677. https://doi.org/10.1038/s41467-018-03098-y · DOI
  6. Fernandez-Casanas M, Chan KL (2018) The Unresolved Problem of DNA Bridging. Genes (Basel) 9. https://doi.org/10.3390/genes9120623 · DOI
  7. Leibowitz ML, Zhang CZ, Pellman D, Chromothripsis (2015) A New Mechanism for Rapid Karyotype Evolution. Annu Rev Genet 49:183–211. https://doi.org/10.1146/annurev-genet-120213-092228 · DOI
  8. Sarlos K et al (2018) Reconstitution of anaphase DNA bridge recognition and disjunction. Nat Struct Mol Biol 25:868–876. https://doi.org/10.1038/s41594-018-0123-8 · DOI
  9. Hengeveld RC et al (2015) Rif1 Is Required for Resolution of Ultrafine DNA Bridges in Anaphase to Ensure Genomic Stability. Dev Cell 34:466–474. https://doi.org/10.1016/j.devcel.2015.06.014 · DOI
  10. Wu L, Hickson ID (2003) The Bloom's syndrome helicase suppresses crossing over during homologous recombination. Nature 426:870–874. https://doi.org/10.1038/nature02253 · DOI
  11. Bussen W, Raynard S, Busygina V, Singh AK, Sung P (2007) Holliday junction processing activity of the BLM-Topo IIIalpha-BLAP75 complex. J Biol Chem 282:31484–31492. https://doi.org/10.1074/jbc.M706116200 · DOI
  12. Wu L et al (2006) BLAP75/RMI1 promotes the BLM-dependent dissolution of homologous recombination intermediates. Proc Natl Acad Sci U S A 103:4068–4073. https://doi.org/10.1073/pnas.0508295103 · DOI
  13. Ellis NA, German J (1996) Molecular genetics of Bloom's syndrome. Hum Mol Genet 5 Spec No, 1457–1463 https://doi.org/10.1093/hmg/5.supplement_1.1457 · DOI
  14. Martin CA et al (2018) Mutations in TOP3A Cause a Bloom Syndrome-like Disorder. Am J Hum Genet 103:221–231. https://doi.org/10.1016/j.ajhg.2018.07.001 · DOI
  15. Bayart E et al (2006) The Bloom syndrome helicase is a substrate of the mitotic Cdc2 kinase. Cell Cycle 5:1681–1686. https://doi.org/10.4161/cc.5.15.3122 · DOI
  16. Leng M et al (2006) MPS1-dependent mitotic BLM phosphorylation is important for chromosome stability. Proc Natl Acad Sci U S A 103:11485–11490. https://doi.org/10.1073/pnas.0601828103 · DOI
  17. Leng M, Besusso D, Jung SY, Wang Y, Qin J (2008) Targeting Plk1 to chromosome arms and regulating chromosome compaction by the PICH ATPase. Cell Cycle 7:1480–1489. https://doi.org/10.4161/cc.7.10.5951 · DOI
  18. Balbo Pogliano C et al (2022) The CDK1-TOPBP1-PLK1 axis regulates the Bloom's syndrome helicase BLM to suppress crossover recombination in somatic cells. Sci Adv 8:eabk0221. https://doi.org/10.1126/sciadv.abk0221 · DOI
  19. Pradhan A, Singh TR, Ali AM, Wahengbam K, Meetei AR (2013) Monopolar spindle 1 (MPS1) protein-dependent phosphorylation of RecQ-mediated genome instability protein 2 (RMI2) at serine 112 is essential for BLM-Topo III alpha-RMI1-RMI2 (BTR) protein complex function upon spindle assembly checkpoint (SAC) activation during mitosis. J Biol Chem 288:33500–33508. https://doi.org/10.1074/jbc.M113.470823 · DOI
  20. Addis Jones O, Tiwari A, Olukoga T, Herbert A, Chan KL (2019) PLK1 facilitates chromosome biorientation by suppressing centromere disintegration driven by BLM-mediated unwinding and spindle pulling. Nat Commun 10:2861. https://doi.org/10.1038/s41467-019-10938-y · DOI
  21. Kong N, Liu Z, Chan YW (2023) RIF1 suppresses the formation of single-stranded ultrafine anaphase bridges via protein phosphatase 1. Cell Rep 42:112032. https://doi.org/10.1016/j.celrep.2023.112032 · DOI
  22. Dutertre S et al (2002) Dephosphorylation and subcellular compartment change of the mitotic Bloom's syndrome DNA helicase in response to ionizing radiation. J Biol Chem 277:6280–6286. https://doi.org/10.1074/jbc.M105735200 · DOI
  23. Kharat SS et al (2016) Mitotic phosphorylation of Bloom helicase at Thr182 is required for its proteasomal degradation and maintenance of chromosomal stability. Oncogene 35:1025–1038. https://doi.org/10.1038/onc.2015.157 · DOI
  24. Lera RF et al (2019) Plk1 protects kinetochore-centromere architecture against microtubule pulling forces. EMBO Rep 20:e48711. https://doi.org/10.15252/embr.201948711 · DOI
  25. Hu P et al (2001) Evidence for BLM and Topoisomerase IIIalpha interaction in genomic stability. Hum Mol Genet 10:1287–1298. https://doi.org/10.1093/hmg/10.12.1287 · DOI
  26. Shorrocks AK et al (2021) The Bloom syndrome complex senses RPA-coated single-stranded DNA to restart stalled replication forks. Nat Commun 12:585. https://doi.org/10.1038/s41467-020-20818-5 · DOI
  27. Hodson C et al (2022) Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability. Proc Natl Acad Sci U S A 119. https://doi.org/10.1073/pnas.2109093119 · DOI
  28. Blackford AN et al (2015) TopBP1 interacts with BLM to maintain genome stability but is dispensable for preventing BLM degradation. Mol Cell 57:1133–1141. https://doi.org/10.1016/j.molcel.2015.02.012 · DOI
  29. Varadi M et al (2022) AlphaFold Protein Structure Database: massively expanding the structural coverage of protein-sequence space with high-accuracy models. Nucleic Acids Res 50:D439–D444. https://doi.org/10.1093/nar/gkab1061 · DOI · Paper page
  30. Singh TR et al (2008) BLAP18/RMI2, a novel OB-fold-containing protein, is an essential component of the Bloom helicase-double Holliday junction dissolvasome. Genes Dev 22:2856–2868. https://doi.org/10.1101/gad.1725108 · DOI
  31. Vassilev LT et al (2006) Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1. Proc Natl Acad Sci U S A 103:10660–10665. https://doi.org/10.1073/pnas.0600447103 · DOI
  32. Choi M et al (2017) TC Mps1 12, a novel Mps1 inhibitor, suppresses the growth of hepatocellular carcinoma cells via the accumulation of chromosomal instability. Br J Pharmacol 174:1810–1825. https://doi.org/10.1111/bph.13782 · DOI
  33. Potapova TA et al (2006) The reversibility of mitotic exit in vertebrate cells. Nature 440:954–958. https://doi.org/10.1038/nature04652 · DOI
  34. Olsen JV et al (2010) Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis. Sci Signal 3:ra3. https://doi.org/10.1126/scisignal.2000475 · DOI
  35. Santamaria A et al (2007) Use of the novel Plk1 inhibitor ZK-thiazolidinone to elucidate functions of Plk1 in early and late stages of mitosis. Mol Biol Cell 18:4024–4036. https://doi.org/10.1091/mbc.e07-05-0517 · DOI
  36. Santamaria A et al (2011) The Plk1-dependent phosphoproteome of the early mitotic spindle. Mol Cell Proteomics 10, M110 004457 https://doi.org/10.1074/mcp.M110.004457 · DOI
  37. Sharma K et al (2014) Ultradeep human phosphoproteome reveals a distinct regulatory nature of Tyr and Ser/Thr-based signaling. Cell Rep 8:1583–1594. https://doi.org/10.1016/j.celrep.2014.07.036 · DOI
  38. Dephoure N et al (2008) A quantitative atlas of mitotic phosphorylation. Proc Natl Acad Sci U S A 105:10762–10767. https://doi.org/10.1073/pnas.0805139105 · DOI · Paper page
  39. Malik R et al (2009) Quantitative analysis of the human spindle phosphoproteome at distinct mitotic stages. J Proteome Res 8:4553–4563. https://doi.org/10.1021/pr9003773 · DOI
  40. Hegemann B et al (2011) Systematic phosphorylation analysis of human mitotic protein complexes. Sci Signal 4:rs12. https://doi.org/10.1126/scisignal.2001993 · DOI
  41. Grosstessner-Hain K et al (2011) Quantitative phospho-proteomics to investigate the polo-like kinase 1-dependent phospho-proteome. Mol Cell Proteomics 10, M111 008540 https://doi.org/10.1074/mcp.M111.008540 · DOI
  42. Kettenbach AN et al (2011) Quantitative phosphoproteomics identifies substrates and functional modules of Aurora and Polo-like kinase activities in mitotic cells. Sci Signal 4:rs5. https://doi.org/10.1126/scisignal.2001497 · DOI
  43. Hiraga SI et al (2017) Human RIF1 and protein phosphatase 1 stimulate DNA replication origin licensing but suppress origin activation. EMBO Rep 18:403–419. https://doi.org/10.15252/embr.201641983 · DOI
  44. Mitsuhashi S et al (2001) Tautomycetin is a novel and specific inhibitor of serine/threonine protein phosphatase type 1, PP1. Biochem Biophys Res Commun 287:328–331. https://doi.org/10.1006/bbrc.2001.5596 · DOI
  45. Kurasawa Y, Yu-Lee LY (2010) PICH and cotargeted Plk1 coordinately maintain prometaphase chromosome arm architecture. Mol Biol Cell 21:1188–1199. https://doi.org/10.1091/mbc.e09-11-0950 · DOI
  46. Orthwein A et al (2014) Mitosis inhibits DNA double-strand break repair to guard against telomere fusions. Science 344:189–193. https://doi.org/10.1126/science.1248024 · DOI
  47. Johnson FB et al (2000) Association of the Bloom syndrome protein with topoisomerase IIIalpha in somatic and meiotic cells. Cancer Res 60:1162–1167
  48. Xu C et al (2015) Accumulation and Phosphorylation of RecQ-Mediated Genome Instability Protein 1 (RMI1) at Serine 284 and Serine 292 during Mitosis. Int J Mol Sci 16:26395–26405. https://doi.org/10.3390/ijms161125965 · DOI
  49. Arnaud L, Pines J, Nigg EA (1998) GFP tagging reveals human Polo-like kinase 1 at the kinetochore/centromere region of mitotic chromosomes. Chromosoma 107:424–429. https://doi.org/10.1007/s004120050326 · DOI
  50. Adams RR et al (2001) Human INCENP colocalizes with the Aurora-B/AIRK2 kinase on chromosomes and is overexpressed in tumour cells. Chromosoma 110:65–74. https://doi.org/10.1007/s004120100130 · DOI
  51. Stucke VM, Sillje HH, Arnaud L, Nigg EA (2002) Human Mps1 kinase is required for the spindle assembly checkpoint but not for centrosome duplication. EMBO J 21:1723–1732. https://doi.org/10.1093/emboj/21.7.1723 · DOI
  52. Shi J et al (2017) A helical bundle in the N-terminal domain of the BLM helicase mediates dimer and potentially hexamer formation. J Biol Chem 292:5909–5920. https://doi.org/10.1074/jbc.M116.761510 · DOI
  53. Kowarz E, Loscher D, Marschalek R (2015) Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol J 10:647–653. https://doi.org/10.1002/biot.201400821 · DOI

Metadata

Authors
Chan, K.-L., Fernández-Casañas, M., Karanika, E., Olukoga, T., Herbert, A., Aliyaskarova, U., & Day, M.
Affiliations
  • University of Sussex (GB)
  • Queen Mary University of London (GB)
ORCID
Venue
Research Square
Publisher
Springer Science and Business Media LLC
Published
2024-05-30
DOI
10.21203/rs.3.rs-4420374/v1
OpenAlex
W4399179049
Cited by
0
References
1
Type
posted content
Language
en
Open access
Licence
Locations
  • Research Square (acceptedVersion, cc-by)
  • Queen Mary Research Online (Queen Mary University of London) (submittedVersion, cc-by)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
Bloom syndrome helicase, BLM, TOP3A, RMI1, RMI2, PICH, CDK1, sister chromatid cohesion, kinetochore, genomic instability, mitosis

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