ROS regulation of RAS and vulva development in Caenorhabditis elegans.
Animals
Animals, Genetically Modified
Caenorhabditis elegans
/ genetics
Caenorhabditis elegans Proteins
/ genetics
Female
Gain of Function Mutation
Gene Expression Regulation, Developmental
Genes, Helminth
/ genetics
Oxidation-Reduction
Oxidoreductases
/ metabolism
Peroxides
/ analysis
Signal Transduction
/ genetics
Transcription Factors
/ metabolism
Vulva
/ growth & development
rac GTP-Binding Proteins
/ metabolism
ras Proteins
/ genetics
Journal
PLoS genetics
ISSN: 1553-7404
Titre abrégé: PLoS Genet
Pays: United States
ID NLM: 101239074
Informations de publication
Date de publication:
06 2020
06 2020
Historique:
received:
20
09
2019
accepted:
07
05
2020
revised:
26
06
2020
pubmed:
17
6
2020
medline:
1
9
2020
entrez:
17
6
2020
Statut:
epublish
Résumé
Reactive oxygen species (ROS) are signalling molecules whose study in intact organisms has been hampered by their potential toxicity. This has prevented a full understanding of their role in organismal processes such as development, aging and disease. In Caenorhabditis elegans, the development of the vulva is regulated by a signalling cascade that includes LET-60ras (homologue of mammalian Ras), MPK-1 (ERK1/2) and LIN-1 (an ETS transcription factor). We show that both mitochondrial and cytoplasmic ROS act on a gain-of-function (gf) mutant of the LET-60ras protein through a redox-sensitive cysteine (C118) previously identified in mammals. We show that the prooxidant paraquat as well as isp-1, nuo-6 and sod-2 mutants, which increase mitochondrial ROS, inhibit the activity of LET-60rasgf on vulval development. In contrast, the antioxidant NAC and loss of sod-1, both of which decrease cytoplasmic H202, enhance the activity of LET-60rasgf. CRISPR replacement of C118 with a non-oxidizable serine (C118S) stimulates LET-60rasgf activity, whereas replacement of C118 with aspartate (C118D), which mimics a strongly oxidised cysteine, inhibits LET-60rasgf. These data strongly suggest that C118 is oxidized by cytoplasmic H202 generated from dismutation of mitochondrial and/or cytoplasmic superoxide, and that this oxidation inhibits LET-60ras. This contrasts with results in cultured mammalian cells where it is mostly nitric oxide, which is not found in worms, that oxidizes C118 and activates Ras. Interestingly, PQ, NAC and the C118S mutation do not act on the phosphorylation of MPK-1, suggesting that oxidation of LET-60ras acts on an as yet uncharacterized MPK-1-independent pathway. We also show that elevated cytoplasmic superoxide promotes vulva formation independently of C118 of LET-60ras and downstream of LIN-1. Finally, we uncover a role for the NADPH oxidases (BLI-3 and DUOX-2) and their redox-sensitive activator CED-10rac in stimulating vulva development. Thus, there are at least three genetically separable pathways by which ROS regulates vulval development.
Identifiants
pubmed: 32544191
doi: 10.1371/journal.pgen.1008838
pii: PGENETICS-D-19-01580
pmc: PMC7319342
doi:
Substances chimiques
CED-10 protein, C elegans
0
Caenorhabditis elegans Proteins
0
Lin-1 protein, C elegans
0
Peroxides
0
Transcription Factors
0
let-60 protein, C elegans
133135-08-7
Bli-3 protein, C elegans
EC 1.-
Oxidoreductases
EC 1.-
rac GTP-Binding Proteins
EC 3.6.5.2
ras Proteins
EC 3.6.5.2
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1008838Subventions
Organisme : CIHR
ID : FDN-159916
Pays : Canada
Organisme : NIH HHS
ID : P40 OD010440
Pays : United States
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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