Genetic recoding to dissect the roles of site-specific protein O-GlcNAcylation

Andrii Gorelik; Sergio Galan Bartual; Vladimir Borodkin; Joby Varghese; Andrew Ferenbach; Daan van Aalten

doi:10.1038/s41594-019-0325-8

Genetic recoding to dissect the roles of site-specific protein O-GlcNAcylation

Andrii Gorelik, Sergio Galan Bartual, Vladimir Borodkin, Joby Varghese, Andrew Ferenbach, Daan van Aalten (Lead / Corresponding author)

Gene Regulation and Expression

Research output: Contribution to journal › Article

4 Citations (Scopus)

33 Downloads (Pure)

Abstract

Modification of specific Ser and Thr residues of nucleocytoplasmic proteins with O-GlcNAc, catalyzed by O-GlcNAc transferase (OGT), is an abundant posttranslational event essential for proper animal development and is dysregulated in various diseases. Due to the rapid concurrent removal by the single O-GlcNAcase (OGA), precise functional dissection of site-specific O-GlcNAc modification in vivo is currently not possible without affecting the entire O-GlcNAc proteome. Exploiting the fortuitous promiscuity of OGT, we show that S-GlcNAc is a hydrolytically stable and accurate structural mimic of O-GlcNAc that can be encoded in mammalian systems with CRISPR-Cas9 in an otherwise unperturbed O-GlcNAcome. Using this approach, we target an elusive Ser 405 O-GlcNAc site on OGA, showing that this site-specific modification affects OGA stability.

Original language	English
Pages (from-to)	1071-1077
Number of pages	7
Journal	Nature Structural & Molecular Biology
Volume	26
Issue number	11
Early online date	6 Nov 2019
DOIs	https://doi.org/10.1038/s41594-019-0325-8
Publication status	Published - Nov 2019

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10.1038/s41594-019-0325-8

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Projects

1 Finished
1 Active

Molecular Mechanisms of O-GICNAC Signalling (Investigator award)

van Aalten, D.

Wellcome Trust

1/03/16 → 31/08/21

Project: Research

Molecular and Cellular Biology 4 Year PhD

Owen-Hughes, T.

Wellcome Trust

1/09/14 → 31/08/18

Project: Research

Cite this

Gorelik, A., Galan Bartual, S., Borodkin, V., Varghese, J., Ferenbach, A., & van Aalten, D. (2019). Genetic recoding to dissect the roles of site-specific protein O-GlcNAcylation. Nature Structural & Molecular Biology, 26(11), 1071-1077. https://doi.org/10.1038/s41594-019-0325-8

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abstract = "Modification of specific Ser and Thr residues of nucleocytoplasmic proteins with O-GlcNAc, catalyzed by O-GlcNAc transferase (OGT), is an abundant posttranslational event essential for proper animal development and is dysregulated in various diseases. Due to the rapid concurrent removal by the single O-GlcNAcase (OGA), precise functional dissection of site-specific O-GlcNAc modification in vivo is currently not possible without affecting the entire O-GlcNAc proteome. Exploiting the fortuitous promiscuity of OGT, we show that S-GlcNAc is a hydrolytically stable and accurate structural mimic of O-GlcNAc that can be encoded in mammalian systems with CRISPR-Cas9 in an otherwise unperturbed O-GlcNAcome. Using this approach, we target an elusive Ser 405 O-GlcNAc site on OGA, showing that this site-specific modification affects OGA stability.",

author = "Andrii Gorelik and {Galan Bartual}, Sergio and Vladimir Borodkin and Joby Varghese and Andrew Ferenbach and {van Aalten}, Daan",

note = "This work was funded by a Wellcome Trust Investigator Award (110061) to DvA and a Wellcome Trust 4-year PhD studentship (105310/Z/14/Z) to AG. ",

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AU - Ferenbach, Andrew

AU - van Aalten, Daan

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N2 - Modification of specific Ser and Thr residues of nucleocytoplasmic proteins with O-GlcNAc, catalyzed by O-GlcNAc transferase (OGT), is an abundant posttranslational event essential for proper animal development and is dysregulated in various diseases. Due to the rapid concurrent removal by the single O-GlcNAcase (OGA), precise functional dissection of site-specific O-GlcNAc modification in vivo is currently not possible without affecting the entire O-GlcNAc proteome. Exploiting the fortuitous promiscuity of OGT, we show that S-GlcNAc is a hydrolytically stable and accurate structural mimic of O-GlcNAc that can be encoded in mammalian systems with CRISPR-Cas9 in an otherwise unperturbed O-GlcNAcome. Using this approach, we target an elusive Ser 405 O-GlcNAc site on OGA, showing that this site-specific modification affects OGA stability.

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