Replication stress, particularly in hard-to-replicate regions such as telomeres and centromeres, leads to the accumulation of replication intermediates that must be processed to ensure proper chromosome segregation. In this study, we identify a critical role for the interaction between RECQ4 and MUS81 in managing such stress. We show that RECQ4 physically interacts with MUS81, targeting it to specific DNA substrates and enhancing its endonuclease activity. Loss of this interaction, results in significant chromosomal segregation defects, including the accumulation of micronuclei, anaphase bridges, and ultrafine bridges (UFBs). Our data further demonstrate that the RECQ4-MUS81 interaction plays an important role in ALT-positive cells, where MUS81 foci primarily colocalise with telomeres, highlighting its role in telomere maintenance. We also observe that a mutation associated with Rothmund-Thomson syndrome, which produces a truncated RECQ4 unable to interact with MUS81, recapitulates these chromosome instability phenotypes. This underscores the importance of RECQ4-MUS81 in safeguarding genome integrity and suggests potential implications for human disease. Our findings demonstrate the RECQ4-MUS81 interaction as a key mechanism in alleviating replication stress at hard-to-replicate regions and highlight its relevance in pathological conditions such as RTS.

Department of Biology Faculty of Medicine Masaryk University Kamenice 5 A7 Brno 62500 Czech Republic

Institute of Biophysics Czech Academy of Sciences Kralovopolska 135 Brno 61200 Czech Republic

National Centre for Biomolecular Research Faculty of Science Masaryk University Kamenice 5 A4 625 00 Brno Czech Republic

Zobrazit více v PubMed

Macheret, M. & Halazonetis, T. D. DNA replication stress as a hallmark of cancer. PubMed DOI

Wassing, I. E. & Esashi, F. RAD51: Beyond the break. PubMed DOI PMC

Li, Z. et al. hDNA2 nuclease/helicase promotes centromeric DNA replication and genome stability. PubMed PMC

Tiwari, A., Addis Jones, O. & Chan, K.-L. 53BP1 can limit sister-chromatid rupture and rearrangements driven by a distinct ultrafine DNA bridging-breakage process. PubMed DOI PMC

Peripolli, S. et al. Oncogenic c-Myc induces replication stress by increasing cohesins chromatin occupancy in a CTCF-dependent manner. PubMed DOI PMC

Segeren, H. A., van Liere, E. A., Riemers, F. M., de Bruin, A. & Westendorp, B. Oncogenic RAS sensitizes cells to drug-induced replication stress via transcriptional silencing of P53. PubMed DOI PMC

Aird, K. M. et al. Suppression of nucleotide metabolism underlies the establishment and maintenance of oncogene-induced senescence. PubMed DOI PMC

Ait Saada, A., Lambert, S. A. E. & Carr, A. M. Preserving replication fork integrity and competence via the homologous recombination pathway. PubMed DOI PMC

Sarbajna, S., Davies, D. & West, S. C. Roles of SLX1-SLX4, MUS81-EME1, and GEN1 in avoiding genome instability and mitotic catastrophe. PubMed DOI PMC

Pepe, A. & West, S. C. MUS81-EME2 promotes replication fork restart. PubMed DOI PMC

Mazina, O. M. & Mazin, A. V. Human Rad54 protein stimulates human Mus81-Eme1 endonuclease. PubMed DOI PMC

Chavdarova, M. et al. Srs2 promotes Mus81-Mms4-mediated resolution of recombination intermediates. PubMed DOI PMC

Murfuni, I. et al. The WRN and MUS81 proteins limit cell death and genome instability following oncogene activation. PubMed DOI

Zhang, R. et al. BLM helicase facilitates Mus81 endonuclease activity in human cells. PubMed DOI

Parvathaneni, S. & Sharma, S. The DNA repair helicase RECQ1 has a checkpoint-dependent role in mediating DNA damage responses induced by gemcitabine. PubMed DOI PMC

Di Marco, S. et al. RECQ5 helicase cooperates with MUS81 endonuclease in processing stalled replication forks at common fragile sites during mitosis. PubMed DOI

Mannuss, A. et al. RAD5A, RECQ4A, and MUS81 have specific functions in homologous recombination and define different pathways of DNA repair in Arabidopsis thaliana. PubMed DOI PMC

Hoki, Y. et al. Growth retardation and skin abnormalities of the Recql4-deficient mouse. PubMed DOI

Kliszczak, M. et al. Interaction of RECQ4 and MCM10 is important for efficient DNA replication origin firing in human cells. PubMed DOI PMC

Sedlackova, H., Cechova, B., Mlcouskova, J. & Krejci, L. RECQ4 selectively recognizes Holliday junctions. PubMed DOI

Kellermayer, R. The versatile RECQL4. PubMed DOI

Zhunussova, G. et al. Mutation spectrum of cancer-associated genes in patients with early onset of colorectal cancer. PubMed DOI PMC

Bakhoum, S. F. & Cantley, L. C. The multifaceted role of chromosomal instability in cancer and its microenvironment. PubMed DOI PMC

Papageorgiou, A. C. et al. Recognition and coacervation of G-quadruplexes by a multifunctional disordered region in RECQ4 helicase. PubMed DOI PMC

Yokoyama, H. et al. Chromosome alignment maintenance requires the MAP RECQL4, mutated in the Rothmund-Thomson syndrome. PubMed PMC

Ke, Y. et al. PICH and BLM limit histone association with anaphase centromeric DNA threads and promote their resolution. PubMed DOI PMC

Zeng, S. et al. Telomere recombination requires the MUS81 endonuclease. PubMed DOI PMC

Scheel, C. et al. Alternative lengthening of telomeres is associated with chromosomal instability in osteosarcomas. PubMed DOI

Wyatt, H. D. M., Sarbajna, S., Matos, J. & West, S. C. Coordinated actions of SLX1-SLX4 and MUS81-EME1 for holliday junction resolution in human cells. PubMed DOI

Chan, Y. W. & West, S. C. Spatial control of the GEN1 Holliday junction resolvase ensures genome stability. PubMed DOI PMC

Xu, X., Chang, C.-W., Li, M., Liu, C. & Liu, Y. Molecular Mechanisms of the RECQ4 Pathogenic Mutations. PubMed DOI PMC

Balraj, P. et al. An unusual mutation in RECQ4 gene leading to Rothmund-Thomson syndrome. PubMed DOI

Chan, Y. W. & West, S. C. A new class of ultrafine anaphase bridges generated by homologous recombination. PubMed DOI PMC

Wilhelm, T., Said, M. & Naim, V. DNA replication stress and chromosomal instability: dangerous liaisons. PubMed PMC

Hengeveld, R. C. C. et al. Rif1 is required for resolution of ultrafine DNA bridges in anaphase to ensure genomic stability. PubMed DOI

Stok, C. et al. FIRRM/C1orf112 is synthetic lethal with PICH and mediates RAD51 dynamics. PubMed DOI

Mann, M. B. et al. Defective sister-chromatid cohesion, aneuploidy and cancer predisposition in a mouse model of type II Rothmund-Thomson syndrome. PubMed DOI

Ghosh, A. K. et al. RECQL4, the protein mutated in Rothmund-Thomson syndrome, functions in telomere maintenance. PubMed DOI PMC

Magidson, V. et al. Adaptive changes in the kinetochore architecture facilitate proper spindle assembly. PubMed DOI PMC

Mankouri, H. W., Huttner, D. & Hickson, I. D. How unfinished business from S-phase affects mitosis and beyond. PubMed DOI PMC

Fang, H. et al. RecQL4-Aurora B kinase axis is essential for cellular proliferation, cell cycle progression, and mitotic integrity. PubMed DOI PMC

Ying, S. et al. MUS81 promotes common fragile site expression. PubMed DOI

Lu, R. & Pickett, H. A. Telomeric replication stress: the beginning and the end for alternative lengthening of telomeres cancers. PubMed DOI PMC

Barefield, C. & Karlseder, J. The BLM helicase contributes to telomere maintenance through processing of late-replicating intermediate structures. PubMed DOI PMC

Sangrithi, M. N. et al. Initiation of DNA replication requires the RECQL4 protein mutated in Rothmund-Thomson syndrome. PubMed DOI

Abe, T. et al. The N-terminal region of RECQL4 lacking the helicase domain is both essential and sufficient for the viability of vertebrate cells. Role of the N-terminal region of RECQL4 in cells. PubMed DOI

Singh, D. K. et al. The human RecQ helicases BLM and RECQL4 cooperate to preserve genome stability. PubMed DOI PMC

Thangavel, S. et al. Human RECQ1 and RECQ4 helicases play distinct roles in DNA replication initiation. PubMed DOI PMC

Matos, J. & West, S. C. Holliday junction resolution: regulation in space and time. PubMed DOI PMC

Matos, J., Blanco, M. G., Maslen, S., Skehel, J. M. & West, S. C. Regulatory control of the resolution of DNA recombination intermediates during meiosis and mitosis. PubMed DOI PMC

Chan, Y. W., Fugger, K. & West, S. C. Unresolved recombination intermediates lead to ultra-fine anaphase bridges, chromosome breaks and aberrations. PubMed DOI PMC

Andrs, M., Hasanova, Z., Oravetzova, A., Dobrovolna, J. & Janscak, P. RECQ5: A mysterious helicase at the interface of DNA replication and transcription. PubMed PMC

Ajeawung, N. F. et al. Mutations in ANAPC1, encoding a scaffold subunit of the anaphase-promoting complex, cause rothmund-thomson syndrome type 1. PubMed DOI PMC

Xu, X. et al. DNA replication initiation factor RECQ4 possesses a role in antagonizing DNA replication initiation. PubMed DOI PMC

Choi, Y.-W. et al. Gene expression profiles in squamous cell cervical carcinoma using array-based comparative genomic hybridization analysis. PubMed DOI

Lu, R. et al. MUS81 participates in the progression of serous ovarian cancer associated with dysfunctional DNA repair system. PubMed DOI PMC

Thul, P. J. & Lindskog, C. The human protein atlas: a spatial map of the human proteome. PubMed DOI PMC

Reinhold, W. C. et al. CellMiner: a web-based suite of genomic and pharmacologic tools to explore transcript and drug patterns in the NCI-60 cell line set. PubMed DOI PMC

Ghodgaonkar, M. M. et al. Phenotypic characterization of missense polymerase-δ mutations using an inducible protein-replacement system. PubMed DOI

Murnane, J. P. Inducible gene expression by DNA rearrangements in human cells. PubMed PMC

Addis Jones, O., Tiwari, A., Olukoga, T., Herbert, A. & Chan, K.-L. PLK1 facilitates chromosome biorientation by suppressing centromere disintegration driven by BLM-mediated unwinding and spindle pulling. PubMed DOI PMC

Sedlackova, H. et al. Equilibrium between nascent and parental MCM proteins protects replicating genomes. PubMed DOI

McQuin, C. et al. CellProfiler 3.0: next-generation image processing for biology. PubMed DOI PMC

Bolte, S. & Cordelières, F. P. A guided tour into subcellular colocalization analysis in light microscopy. PubMed DOI

Notredame, C., Higgins, D. G. & Heringa, J. T-Coffee: A novel method for fast and accurate multiple sequence alignment. PubMed DOI

Robert, X. & Gouet, P. Deciphering key features in protein structures with the new ENDscript server. PubMed DOI PMC

Jumper, J. et al. Highly accurate protein structure prediction with AlphaFold. PubMed DOI PMC

Pettersen, E. F. et al. UCSF chimera-a visualization system for exploratory research and analysis. PubMed DOI

Macris, M. A., Krejci, L., Bussen, W., Shimamoto, A. & Sung, P. Biochemical characterization of the RECQ4 protein, mutated in Rothmund-Thomson syndrome. PubMed DOI

Garcia, P. L., Liu, Y., Jiricny, J., West, S. C. & Janscak, P. Human RECQ5beta, a protein with DNA helicase and strand-annealing activities in a single polypeptide. PubMed DOI PMC