Nse5/6 is a negative regulator of the ATPase activity of the Smc5/6 complex.
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Date
2021-05-07Author
Hallett, ST
Schellenberger, P
Zhou, L
Beuron, F
Morris, E
Murray, JM
Oliver, AW
Type
Journal Article
Metadata
Show full item recordAbstract
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.
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Subject
Escherichia coli
Saccharomyces cerevisiae
Cell Cycle Proteins
Saccharomyces cerevisiae Proteins
Chromosomal Proteins, Non-Histone
DNA
Microscopy, Electron, Transmission
Adenosine Triphosphatases
Research team
Structural Electron Microscopy
Structural Electron Microscopy
Language
eng
Date accepted
2021-03-22
Citation
Nucleic acids research, 2021, 49 (8), pp. 4534 - 4549
Publisher
OXFORD UNIV PRESS