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

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posted on 2023-06-09, 23:32 authored by Stephen Hallett, Pascale Schellenberger, Lihong Zhou, Fabienne Beuron, Ed Morris, Jo Murray, Antony OliverAntony Oliver
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.

Funding

How do Smc5/6 interactions with DNA coordinate replication and recombination?; G2119; MRC-MEDICAL RESEARCH COUNCIL; MR/P018955/1

History

Publication status

  • Published

File Version

  • Published version

Journal

Nucleic Acids Research

ISSN

0305-1048

Publisher

Oxford University Press

Issue

8

Volume

49

Page range

4534-4549

Department affiliated with

  • Sussex Centre for Genome Damage Stability Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2021-04-09

First Open Access (FOA) Date

2021-04-21

First Compliant Deposit (FCD) Date

2021-04-21

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