TY - JOUR
T1 - Genetic recoding to dissect the roles of site-specific protein O-GlcNAcylation
AU - Gorelik, Andrii
AU - Galan Bartual, Sergio
AU - Borodkin, Vladimir
AU - Varghese, Joby
AU - Ferenbach, Andrew
AU - van Aalten, Daan
N1 - 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.
PY - 2019/11
Y1 - 2019/11
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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85075090236&partnerID=8YFLogxK
U2 - 10.1038/s41594-019-0325-8
DO - 10.1038/s41594-019-0325-8
M3 - Article
C2 - 31695185
VL - 26
SP - 1071
EP - 1077
JO - Nature Structural & Molecular Biology
JF - Nature Structural & Molecular Biology
SN - 1545-9993
IS - 11
ER -