Nse5/6 is a negative regulator of the ATPase activity of the Smc5/6 complex

Hallett, Stephen T, Schellenberger, Pascale, Zhou, Lihong, Beuron, Fabienne, Morris, Ed, Murray, Johanne M and Oliver, Antony W (2021) Nse5/6 is a negative regulator of the ATPase activity of the Smc5/6 complex. Nucleic Acids Research, 49 (8). pp. 4534-4549. ISSN 0305-1048

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The multi-component Smc5/6 complex plays a critical role in the resolution of recombination intermediates formed during mitosis and meiosis, and in the cellular response to replication stress. Using recombinant proteins, we have reconstituted a series of defined Saccharomyces cerevisiae Smc5/6 complexes, visualised them by negative stain electron microscopy, and tested their ability to function as an ATPase. We find that only the six protein ‘holo-complex’ is capable of turning over ATP and that its activity is significantly increased by the addition of double-stranded DNA to reaction mixes. Furthermore, stimulation is wholly dependent on functional ATP-binding pockets in both Smc5 and Smc6. Importantly, we demonstrate that budding yeast Nse5/6 acts as a negative regulator of Smc5/6 ATPase activity, binding to the head-end of the complex to suppress turnover, irrespective of the DNA-bound status of the complex.

Item Type: Article
Keywords: DNA Repair, Electron Microscopy, Structural Biology, Structural Maintenance of Chromosomes
Schools and Departments: School of Life Sciences > Sussex Centre for Genome Damage and Stability
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 09 Apr 2021 07:03
Last Modified: 28 Feb 2022 14:45
URI: http://sro.sussex.ac.uk/id/eprint/98318

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Project NameSussex Project NumberFunderFunder Ref
How do Smc5/6 interactions with DNA coordinate replication and recombination?G2119MRC-MEDICAL RESEARCH COUNCILMR/P018955/1