Nse5/6 is a negative regulator of the ATPase activity of the Smc5/6 complex.
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Embargo End Date
ICR Authors
Authors
Hallett, ST
Schellenberger, P
Zhou, L
Beuron, F
Morris, E
Murray, JM
Oliver, AW
Schellenberger, P
Zhou, L
Beuron, F
Morris, E
Murray, JM
Oliver, AW
Document Type
Journal Article
Date
2021-05-07
Date Accepted
2021-03-22
Abstract
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.
Citation
Nucleic acids research, 2021, 49 (8), pp. 4534 - 4549
Source Title
Publisher
OXFORD UNIV PRESS
ISSN
0305-1048
eISSN
1362-4962
Collections
Research Team
Structural Electron Microscopy
Structural Electron Microscopy
Structural Electron Microscopy