title: Identification of nuclear target proteins for S-nitrosylation in pathogen-treated Arabidopsis thaliana cell cultures.
abstract: Nitric oxide (NO) is a significant signalling molecule involved in the regulation of many different physiological processes in plants. One of the most imperative regulatory modes of action of NO is protein S-nitrosylation--the covalent attachment of an NO group to the sulfur atom of cysteine residues. In this study, we focus on S-nitrosylation of Arabidopsis nuclear proteins after pathogen infection. After treatment of Arabidopsis suspension cell cultures with pathogens, nuclear proteins were extracted and treated with the S-nitrosylating agent S-nitrosoglutathione (GSNO). A biotin switch assay was performed and biotin-labelled proteins were purified by neutravidin affinity chromatography and identified by mass spectrometry. A total of 135 proteins were identified, whereas nuclear localization has been described for 122 proteins of them. 117 of these proteins contain at least one cysteine residue. Most of the S-nitrosylated candidates were involved in protein and RNA metabolism, stress response, and cell organization and division. Interestingly, two plant-specific histone deacetylases were identified suggesting that nitric oxide regulated epigenetic processes in plants. In sum, this work provides a new collection of targets for protein S-nitrosylation in Arabidopsis and gives insight into the regulatory function of NO in the nucleus during plant defense response. Moreover, our data extend the knowledge on the regulatory function of NO in events located in the nucleus.
authors.affiliation: Institute of Biochemical Plant Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
authors.forename: Mounira
authors.initials: M
authors.lastname: Chaki
authors.affiliation: Institute of Biochemical Plant Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
authors.forename: Azam
authors.initials: A
authors.lastname: Shekariesfahlan
authors.affiliation: Institute of Biochemical Plant Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Ageeva
authors.affiliation: Institute of Biochemical Plant Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Mengel
authors.affiliation: Research Unit Protein Science, Helmholtz Zentrum München-German Research Center for Environmental Health, 85764 Neuherberg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: von Toerne
authors.affiliation: Institute of Biochemical Plant Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany; Chair of Biochemical Plant Pathology, Technische Universität München, 85354 Freising, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Durner
authors.affiliation: Institute of Biochemical Plant Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany. Electronic address: lindermayr@helmholtz-muenchen.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Lindermayr
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000166:cytology / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D003545:Cysteine / Q000378:metabolism; D015151:Immunoblotting; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000096:biosynthesis; D015538:Nitrosation; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010936:Plant Extracts / Q000378:metabolism; D044224:Pseudomonas syringae / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D012984:Software
chemical_list: D029681:Arabidopsis Proteins; D009687:Nuclear Proteins; D010936:Plant Extracts; D011994:Recombinant Proteins; D009569:Nitric Oxide; D003545:Cysteine
keywords: Biotin switch technique; Cysteine residues; Nitric oxide; Plant defense response; Post-translational modification; Protein S-nitrosylation