Redox requirements for ubiquitin-like urmylation of Ahp1, a 2-Cys peroxiredoxin from yeast.
Date
2020-02-01Author
Brachmann, C
Kaduhr, L
Jüdes, A
Ravichandran, KE
West, JD
Glatt, S
Schaffrath, R
Type
Journal Article
Metadata
Show full item recordAbstract
The yeast peroxiredoxin Ahp1, like related anti-oxidant enzymes in other species, undergoes urmylation, a lysine-directed conjugation to ubiquitin-like modifier Urm1. Ahp1 assembles into a homodimer that detoxifies peroxides via forming intersubunit disulfides between peroxidatic and resolving cysteines that are subsequently reduced by the thioredoxin system. Although urmylation coincides with oxidative stress, it is unclear how this modification happens on a molecular level and whether it affects peroxiredoxin activity. Here, we report that thioredoxin mutants decrease Ahp1 urmylation in yeast and each subunit of the oxidized Ahp1 dimer is modified by Urm1 suggesting coupling of urmylation to dimerization. Consistently, Ahp1 mutants unable to form dimers, fail to be urmylated as do mutants that lack the peroxidatic cysteine. Moreover, Ahp1 urmylation involves at least two lysine residues close to the catalytic cysteines and can be prevented in yeast cells exposed to high organic peroxide concentrations. Our results elucidate redox requirements and molecular determinants critical for Ahp1 urmylation, thus providing insights into a potential link between oxidant defense and Urm1 utilization in cells.
Collections
Subject
2-Cys peroxiredoxin
Ahp1
Dimer interface
Protein urmylation
Redox-active thiols
Saccharomyces cerevisiae
Thioredoxin system
Ubiquitin-related modifier Urm1
Catalytic Domain
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Fungal
Models, Molecular
Mutation
Oxidation-Reduction
Peroxides
Peroxiredoxins
Protein Conformation
Protein Multimerization
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Language
eng
Date accepted
2020-01-17
License start date
2020-02-01
Citation
Redox Biology, 2020, 30 pp. 101438 -
Publisher
ELSEVIER