Proteolysis of HCF-1 by Ser/Thr glycosylation-incompetent O-GlcNAc transferase

UDP-GlcNAc complexes

Vaibhav Kapuria, Ute F. Röhrig, Tanja Bhuiyan, Vladimir S. Borodkin, Daan M F van Aalten, Vincent Zoete, Winship Herr (Lead / Corresponding author)

Research output: Contribution to journalArticle

8 Citations (Scopus)
79 Downloads (Pure)

Abstract

In complex with the cosubstrate UDP-N-acetylglucosamine (UDP-GlcNAc), O-linked-GlcNAc transferase (OGT) catalyzes Ser/Thr O-GlcNAcylation of many cellular proteins and proteolysis of the transcriptional coregulator HCF-1. Such a dual glycosyltransferase-protease activity, which occurs in the same active site, is unprecedented and integrates both reversible and irreversible forms of protein post-translational modification within one enzyme. Although occurring within the same active site, we show here that glycosylation and proteolysis occur through separable mechanisms. OGT consists of tetratricopeptide repeat (TPR) and catalytic domains, which, together with UDP-GlcNAc, are required for both glycosylation and proteolysis. Nevertheless, a specific TPR domain contact with the HCF-1 substrate is critical for proteolysis but not Ser/Thr glycosylation. In contrast, key catalytic domain residues and even a UDP-GlcNAc oxygen important for Ser/Thr glycosylation are irrelevant for proteolysis. Thus, from a dual glycosyltransferase-protease, essentially single-activity enzymes can be engineered both in vitro and in vivo. Curiously, whereas OGT-mediated HCF-1 proteolysis is limited to vertebrate species, invertebrate OGTs can cleave human HCF-1. We present a model for the evolution of HCF-1 proteolysis by OGT.

Original languageEnglish
Pages (from-to)960-972
Number of pages13
JournalGenes and Development
Volume30
Issue number8
Early online date7 Apr 2016
DOIs
Publication statusPublished - 15 Apr 2016

Fingerprint

Uridine Diphosphate
Glycosylation
Proteolysis
Catalytic Domain
Glycosyltransferases
Peptide Hydrolases
Uridine Diphosphate N-Acetylglucosamine
Enzymes
Invertebrates
Post Translational Protein Processing
O-GlcNAc transferase
Vertebrates
Oxygen

Keywords

  • O-GlcNAcylation
  • Post-translation modifications
  • Proteolysis

Cite this

Kapuria, Vaibhav ; Röhrig, Ute F. ; Bhuiyan, Tanja ; Borodkin, Vladimir S. ; van Aalten, Daan M F ; Zoete, Vincent ; Herr, Winship. / Proteolysis of HCF-1 by Ser/Thr glycosylation-incompetent O-GlcNAc transferase : UDP-GlcNAc complexes. In: Genes and Development. 2016 ; Vol. 30, No. 8. pp. 960-972.
@article{1f8b8a6772014817ba2d6cf9db03e18b,
title = "Proteolysis of HCF-1 by Ser/Thr glycosylation-incompetent O-GlcNAc transferase: UDP-GlcNAc complexes",
abstract = "In complex with the cosubstrate UDP-N-acetylglucosamine (UDP-GlcNAc), O-linked-GlcNAc transferase (OGT) catalyzes Ser/Thr O-GlcNAcylation of many cellular proteins and proteolysis of the transcriptional coregulator HCF-1. Such a dual glycosyltransferase-protease activity, which occurs in the same active site, is unprecedented and integrates both reversible and irreversible forms of protein post-translational modification within one enzyme. Although occurring within the same active site, we show here that glycosylation and proteolysis occur through separable mechanisms. OGT consists of tetratricopeptide repeat (TPR) and catalytic domains, which, together with UDP-GlcNAc, are required for both glycosylation and proteolysis. Nevertheless, a specific TPR domain contact with the HCF-1 substrate is critical for proteolysis but not Ser/Thr glycosylation. In contrast, key catalytic domain residues and even a UDP-GlcNAc oxygen important for Ser/Thr glycosylation are irrelevant for proteolysis. Thus, from a dual glycosyltransferase-protease, essentially single-activity enzymes can be engineered both in vitro and in vivo. Curiously, whereas OGT-mediated HCF-1 proteolysis is limited to vertebrate species, invertebrate OGTs can cleave human HCF-1. We present a model for the evolution of HCF-1 proteolysis by OGT.",
keywords = "O-GlcNAcylation, Post-translation modifications, Proteolysis",
author = "Vaibhav Kapuria and R{\"o}hrig, {Ute F.} and Tanja Bhuiyan and Borodkin, {Vladimir S.} and {van Aalten}, {Daan M F} and Vincent Zoete and Winship Herr",
year = "2016",
month = "4",
day = "15",
doi = "10.1101/gad.275925.115",
language = "English",
volume = "30",
pages = "960--972",
journal = "Genes & Development",
issn = "0890-9369",
publisher = "Cold Spring Harbor Laboratory Press",
number = "8",

}

Proteolysis of HCF-1 by Ser/Thr glycosylation-incompetent O-GlcNAc transferase : UDP-GlcNAc complexes. / Kapuria, Vaibhav; Röhrig, Ute F.; Bhuiyan, Tanja; Borodkin, Vladimir S.; van Aalten, Daan M F; Zoete, Vincent; Herr, Winship (Lead / Corresponding author).

In: Genes and Development, Vol. 30, No. 8, 15.04.2016, p. 960-972.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Proteolysis of HCF-1 by Ser/Thr glycosylation-incompetent O-GlcNAc transferase

T2 - UDP-GlcNAc complexes

AU - Kapuria, Vaibhav

AU - Röhrig, Ute F.

AU - Bhuiyan, Tanja

AU - Borodkin, Vladimir S.

AU - van Aalten, Daan M F

AU - Zoete, Vincent

AU - Herr, Winship

PY - 2016/4/15

Y1 - 2016/4/15

N2 - In complex with the cosubstrate UDP-N-acetylglucosamine (UDP-GlcNAc), O-linked-GlcNAc transferase (OGT) catalyzes Ser/Thr O-GlcNAcylation of many cellular proteins and proteolysis of the transcriptional coregulator HCF-1. Such a dual glycosyltransferase-protease activity, which occurs in the same active site, is unprecedented and integrates both reversible and irreversible forms of protein post-translational modification within one enzyme. Although occurring within the same active site, we show here that glycosylation and proteolysis occur through separable mechanisms. OGT consists of tetratricopeptide repeat (TPR) and catalytic domains, which, together with UDP-GlcNAc, are required for both glycosylation and proteolysis. Nevertheless, a specific TPR domain contact with the HCF-1 substrate is critical for proteolysis but not Ser/Thr glycosylation. In contrast, key catalytic domain residues and even a UDP-GlcNAc oxygen important for Ser/Thr glycosylation are irrelevant for proteolysis. Thus, from a dual glycosyltransferase-protease, essentially single-activity enzymes can be engineered both in vitro and in vivo. Curiously, whereas OGT-mediated HCF-1 proteolysis is limited to vertebrate species, invertebrate OGTs can cleave human HCF-1. We present a model for the evolution of HCF-1 proteolysis by OGT.

AB - In complex with the cosubstrate UDP-N-acetylglucosamine (UDP-GlcNAc), O-linked-GlcNAc transferase (OGT) catalyzes Ser/Thr O-GlcNAcylation of many cellular proteins and proteolysis of the transcriptional coregulator HCF-1. Such a dual glycosyltransferase-protease activity, which occurs in the same active site, is unprecedented and integrates both reversible and irreversible forms of protein post-translational modification within one enzyme. Although occurring within the same active site, we show here that glycosylation and proteolysis occur through separable mechanisms. OGT consists of tetratricopeptide repeat (TPR) and catalytic domains, which, together with UDP-GlcNAc, are required for both glycosylation and proteolysis. Nevertheless, a specific TPR domain contact with the HCF-1 substrate is critical for proteolysis but not Ser/Thr glycosylation. In contrast, key catalytic domain residues and even a UDP-GlcNAc oxygen important for Ser/Thr glycosylation are irrelevant for proteolysis. Thus, from a dual glycosyltransferase-protease, essentially single-activity enzymes can be engineered both in vitro and in vivo. Curiously, whereas OGT-mediated HCF-1 proteolysis is limited to vertebrate species, invertebrate OGTs can cleave human HCF-1. We present a model for the evolution of HCF-1 proteolysis by OGT.

KW - O-GlcNAcylation

KW - Post-translation modifications

KW - Proteolysis

UR - http://www.scopus.com/inward/record.url?scp=84963852308&partnerID=8YFLogxK

U2 - 10.1101/gad.275925.115

DO - 10.1101/gad.275925.115

M3 - Article

VL - 30

SP - 960

EP - 972

JO - Genes & Development

JF - Genes & Development

SN - 0890-9369

IS - 8

ER -