title: Glutaredoxin regulates vascular development by reversible glutathionylation of sirtuin 1.
abstract: Embryonic development depends on complex and precisely orchestrated signaling pathways including specific reduction/oxidation cascades. Oxidoreductases of the thioredoxin family are key players conveying redox signals through reversible posttranslational modifications of protein thiols. The importance of this protein family during embryogenesis has recently been exemplified for glutaredoxin 2, a vertebrate-specific glutathione-disulfide oxidoreductase with a critical role for embryonic brain development. Here, we discovered an essential function of glutaredoxin 2 during vascular development. Confocal microscopy and time-lapse studies based on two-photon microscopy revealed that morpholino-based knockdown of glutaredoxin 2 in zebrafish, a model organism to study vertebrate embryogenesis, resulted in a delayed and disordered blood vessel network. We were able to show that formation of a functional vascular system requires glutaredoxin 2-dependent reversible S-glutathionylation of the NAD(+)-dependent protein deacetylase sirtuin 1. Using mass spectrometry, we identified a cysteine residue in the conserved catalytic region of sirtuin 1 as target for glutaredoxin 2-specific deglutathionylation. Thereby, glutaredoxin 2-mediated redox regulation controls enzymatic activity of sirtuin 1, a mechanism we found to be conserved between zebrafish and humans. These results link S-glutathionylation to vertebrate development and successful embryonic angiogenesis.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Department of Molecular Tumor and Cell Biology, and Institute of Environmental Medicine, Karolinska Institutet, 17177, Stockholm, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Bräutigam
authors.forename: Lasse Dahl Ejby
authors.initials: LD
authors.lastname: Jensen
authors.forename: Gereon
authors.initials: G
authors.lastname: Poschmann
authors.forename: Staffan
authors.initials: S
authors.lastname: Nyström
authors.forename: Sarah
authors.initials: S
authors.lastname: Bannenberg
authors.forename: Kristian
authors.initials: K
authors.lastname: Dreij
authors.forename: Klaudia
authors.initials: K
authors.lastname: Lepka
authors.forename: Timour
authors.initials: T
authors.lastname: Prozorovski
authors.forename: Sergio J
authors.initials: SJ
authors.lastname: Montano
authors.forename: Orhan
authors.initials: O
authors.lastname: Aktas
authors.forename: Per
authors.initials: P
authors.lastname: Uhlén
authors.forename: Kai
authors.initials: K
authors.lastname: Stühler
authors.forename: Yihai
authors.initials: Y
authors.lastname: Cao
authors.forename: Arne
authors.initials: A
authors.lastname: Holmgren
authors.forename: Carsten
authors.initials: C
authors.lastname: Berndt
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002319:Cardiovascular System / Q000196:embryology*; D017931:DNA Primers / Q000235:genetics; D055785:Gene Knockdown Techniques; D054477:Glutaredoxins / Q000235:genetics / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010084:Oxidation-Reduction; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D056564:Sirtuin 1 / Q000378:metabolism*; D059008:Time-Lapse Imaging; D015027:Zebrafish
chemical_list: D017931:DNA Primers; D054477:Glutaredoxins; D056564:Sirtuin 1; D005978:Glutathione
keywords: cardiovascular system; proteomics