Journal article
Mechanism of Bloom syndrome complex assembly required for double Holliday junction dissolution and genome stability
C Hodson, JKK Low, S van Twest, SE Jones, P Swuec, V Murphy, K Tsukada, M Fawkes, R Bythell-Douglas, A Davies, JK Holien, JJ O'Rourke, BL Parker, A Glaser, MW Parker, JP Mackay, AN Blackford, A Costa, AJ Deans
Proceedings of the National Academy of Sciences of the United States of America | NATL ACAD SCIENCES | Published : 2022
Abstract
The RecQ-like helicase BLM cooperates with topoisomerase IIIα, RMI1, and RMI2 in a heterotetrameric complex (the “Bloom syndrome complex”) for dissolution of double Holliday junctions, key intermediates in homologous recombination. Mutations in any component of the Bloom syndrome complex can cause genome instability and a highly cancer-prone disorder called Bloom syndrome. Some heterozygous carriers are also predisposed to breast cancer. To understand how the activities of BLM helicase and topoisomerase IIIα are coupled, we purified the active four-subunit complex. Chemical cross-linking and mass spectrometry revealed a unique architecture that links the helicase and topoisomerase domains. U..
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Awarded by University of Sydney
Funding Acknowledgements
We thank Tao-Shih Hsieh, Daniela Stock, and Ross Chapman for providing plasmids; Steve West for providing plasmids and GEN1 antibody; Timothy Richmond for providing the Multibac system; and Ian Hickson for RMI2 and TopoIIIa antibodies. Additionally, we thank Nicola O'Reilly (The Francis Crick Institute Peptide Chemistry Platform) for preparing the fluorescent peptides and the peptide arrays and Steve West and James Berger for providing the initial discussions and impetus to start this project. Mass spectrometry was facilitated by access to Sydney Mass Spectrometry, a core research facility at the University of Sydney. This work was funded by The Cancer Council of Victoria (A.C. and A.J. D.); National Health and Medical Research Council Australia Grants GNT1033592 (to A.J.D.), GNT1181110 (to A.J.D.), GNT1126357 (to J.P.M.), and GNT1146355 (to J.P.M.); The Francis Crick Institute (which receives its core funding from Cancer Research UK, the UK Medical Research Council, and the Wellcome Trust); and the Victorian Government's OIS Program. K.T. is a Japan Society for the Promotion of Science Postdoctoral Research Fellow (JP20J13601). J.K.H. is an RMIT Vice Chancellor's Research Fellow and was a joint Cure Cancer/Leukaemia Foundation Australia Postdoctoral Fellow. A.N.B. is a Cancer Research UK Career Development Fellow (C29215/A20772) and an Against Breast Cancer/Oriel College Research Fellow.