The conserved threonine-rich region of the HCF-1<sub>PRO</sub> repeat activates promiscuous OGT:UDP-GlcNAc glycosylation and proteolysis activities.

Détails

Ressource 1Télécharger: J. Biol. Chem.-2018-Kapuria-17754-68.pdf (3419.87 [Ko])
Etat: Public
Version: Final published version
ID Serval
serval:BIB_2C5B11AB86FD
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
The conserved threonine-rich region of the HCF-1<sub>PRO</sub> repeat activates promiscuous OGT:UDP-GlcNAc glycosylation and proteolysis activities.
Périodique
The Journal of biological chemistry
Auteur(s)
Kapuria V., Röhrig U.F., Waridel P., Lammers F., Borodkin V.S., van Aalten DMF, Zoete V., Herr W.
ISSN
1083-351X (Electronic)
ISSN-L
0021-9258
Statut éditorial
Publié
Date de publication
16/11/2018
Peer-reviewed
Oui
Volume
293
Numéro
46
Pages
17754-17768
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
O-Linked GlcNAc transferase (OGT) possesses dual glycosyltransferase-protease activities. OGT thereby stably glycosylates serines and threonines of numerous proteins and, via a transient glutamate glycosylation, cleaves a single known substrate-the so-called HCF-1 <sub>PRO</sub> repeat of the transcriptional co-regulator host-cell factor 1 (HCF-1). Here, we probed the relationship between these distinct glycosylation and proteolytic activities. For proteolysis, the HCF-1 <sub>PRO</sub> repeat possesses an important extended threonine-rich region that is tightly bound by the OGT tetratricopeptide-repeat (TPR) region. We report that linkage of this HCF-1 <sub>PRO</sub> -repeat, threonine-rich region to heterologous substrate sequences also potentiates robust serine glycosylation with the otherwise poor R <sub>p</sub> -αS-UDP-GlcNAc diastereomer phosphorothioate and UDP-5S-GlcNAc OGT co-substrates. Furthermore, it potentiated proteolysis of a non-HCF-1 <sub>PRO</sub> -repeat cleavage sequence, provided it contained an appropriately positioned glutamate residue. Using serine- or glutamate-containing HCF-1 <sub>PRO</sub> -repeat sequences, we show that proposed OGT-based or UDP-GlcNAc-based serine-acceptor residue activation mechanisms can be circumvented independently, but not when disrupted together. In contrast, disruption of both proposed activation mechanisms even in combination did not inhibit OGT-mediated proteolysis. These results reveal a multiplicity of OGT glycosylation strategies, some leading to proteolysis, which could be targets of alternative molecular regulatory strategies.
Mots-clé
Amino Acid Motifs, Amino Acid Sequence, Endopeptidases/genetics, Endopeptidases/metabolism, Glycosylation, Host Cell Factor C1/genetics, Host Cell Factor C1/metabolism, Humans, Molecular Dynamics Simulation, Multifunctional Enzymes/genetics, Multifunctional Enzymes/metabolism, Mutation, N-Acetylglucosaminyltransferases/genetics, N-Acetylglucosaminyltransferases/metabolism, Proteolysis, Stereoisomerism, Substrate Specificity, Uridine Diphosphate N-Acetylglucosamine/analogs & derivatives, Uridine Diphosphate N-Acetylglucosamine/metabolism, O-GlcNAcylation, O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT), enzyme mechanism, glycobiology, host-cell factor-1, post-translational modification (PTM)
Pubmed
Web of science
Open Access
Oui
Création de la notice
27/11/2018 10:36
Dernière modification de la notice
20/08/2019 14:11
Données d'usage