Characterization of RBPome in Oxidative Stress Conditions.
In vivo UV crosslinking
Mass spectrometry
Oxidative stress
Post-transcriptional gene regulation
Proteomics
RNA-binding proteins
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
3
6
2022
pubmed:
4
6
2022
medline:
9
6
2022
Statut:
ppublish
Résumé
Cellular redox signaling is triggered by accumulation of various reactive oxygen species (ROS) that integrate with other signaling cascades to enable plants to ultimately respond to (a)biotic stresses. The identification of key regulators underlying redox signaling networks is therefore of high priority. This chapter describes an improved mRNA interactome capture method that allows to systematically detect oxidative stress responsive regulators in the post-transcriptional gene regulation (PTGR) pathway. The protocol includes PSB-D suspension cell culture preparation, setup of oxidative stress conditions, short-term exposure to UV irradiation, cell lysis, pull-down and purification of crosslinked messenger ribonucleoproteins, their mass spectrometric analyses, and identification of proteome by statistical analyses. As result, a comprehensive inventory of the functional oxidative stress responsive RBPome (OxRBPome) is generated, which paves the way toward new insights into PTGR processes in redox signaling.
Identifiants
pubmed: 35657526
doi: 10.1007/978-1-0716-2469-2_19
doi:
Substances chimiques
Proteome
0
Reactive Oxygen Species
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
259-275Informations de copyright
© 2022. Springer Science+Business Media, LLC, part of Springer Nature.
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