Abstract
The Endosomal Sorting Complex Required for Transport (ESCRT) pathway, through inverse topology membrane remodeling, is involved in many biological functions, such as ubiquitinated membrane receptor trafficking and degradation, multivesicular bodies (MVB) formation and cytokinesis. Dysfunctions in ESCRT pathway have been associated to several human pathologies, such as cancers and neurodegenerative diseases. The ESCRT machinery is also hijacked by many enveloped viruses to bud away from the plasma membrane of infected cells. Human tumor susceptibility gene 101 (Tsg101) protein is an important ESCRT-I complex component. The structure of the N-terminal ubiquitin E2 variant (UEV) domain of Tsg101 (Tsg101-UEV) comprises an ubiquitin binding pocket next to a late domain [P(S/T)AP] binding groove. These two binding sites have been shown to be involved both in the physiological roles of ESCRT-I and in the release of the viral particles, and thus are attractive targets for antivirals. The structure of the Tsg101-UEV domain has been characterized, using X-ray crystallography or NMR spectroscopy, either in its apo-state or bound to ubiquitin or late domains. In this study, we report the backbone NMR resonance assignments, including the proline signals, of the apo human Tsg101-UEV domain, that so far was not publicly available. These data, that are in good agreement with the crystallographic structure of Tsg101-UEV domain, can therefore be used for further NMR studies, including protein-protein interaction studies and drug discovery.
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Data availability
All the chemical shift values of apo human Tsg101-UEV domain were deposited in the Biological Magnetic Resonance Data Bank (BMRB) under accession code 50765.
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Acknowledgements
The NMR facilities were funded by the Nord Region Council, CNRS, Institut Pasteur de Lille, European Union (FEDER), French Research Ministry and University of Lille. Financial support from the NMR division of Infranalytics (FR 2054 CNRS) is gratefully acknowledged.
Funding
This study was supported by the French National Agency for Research on AIDS and Viral Hepatitis (ANRS) Grant ECTZ101316, and a PhD fellowship from ANRS to D.M. (ECTZ103422).
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D.M. and X.H performed protein expression and purification. F-X.C. and E.B. collected the NMR data. D.M. analysed the data and wrote the first draft of the manuscript. D.M. prepared Figs. 1, 2 and 3. All authors commented on previous versions of the manuscript. All authors reviewed the manuscript. Funding acquisition and supervision were done by X.H.
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Moschidi, D., Cantrelle, FX., Boll, E. et al. Backbone NMR resonance assignment of the apo human Tsg101-UEV domain. Biomol NMR Assign 17, 49–54 (2023). https://doi.org/10.1007/s12104-023-10119-5
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DOI: https://doi.org/10.1007/s12104-023-10119-5