Analysis of oligomeric protein complexes in the chloroplast sub-proteome of nucleic acid-binding proteins from mustard reveals potential redox regulators of plastid gene expression

verfasst von: Yvonne Schröter, Sebastian Steiner, Kevin Matthäi, Thomas Pfannschmidt
Abstract: Photosynthetic light quality acclimation in plants involves redox-controlled changes in plastid gene expression. To study proteins potentially involved in this regulation, we isolated low-abundant chloroplast nucleic acid-binding proteins from the crucifere mustard (Sinapis alba) and investigated if photosynthetic redox signals affect their composition and/or oligomeric structure. We purified chloroplasts from plants subjected to light quality shifts and applied organelle lysates to heparin-Sepharose chromatography followed by 2-D blue native PAGE. We studied accumulation and structure of oligomeric protein complexes and applied MS/MS to identify them. We found ten oligomeric protein complexes of higher order and eleven smaller protein complexes or spots including plastid-encoded RNA polymerase (PEP), plastid transcriptionally active chromosome proteins, RNA-binding proteins, ribosomal subunits and chaperones. A translation elongation factor was found to be the only protein displaying major differences in its amounts in response to the growth lights. Furthermore, we found a novel thioredoxin as a subunit of the PEP, a 2-Cys-peroxiredoxin complex and a (soluble) ferredoxin:NADP-oxido-reductase, which represent potential redox regulator of plastid gene expression. A T-DNA knock-out line of the thioredoxin from Arabidopsis exhibits a yellowish-pale phenotype, demonstrating that this novel PEP subunit is essential for proper plastid development.
Organisationseinheit(en): Institut für Botanik
Externe Organisation(en): Friedrich-Schiller-Universität Jena
Typ: Artikel
Journal: PROTEOMICS
Band: 10
Seiten: 2191-204
Anzahl der Seiten: 14
ISSN: 1615-9853
Publikationsdatum: 06.2010
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
Elektronische Version(en): https://doi.org/10.1002/pmic.200900678 (Zugang: Unbekannt)

BibTeX

@article{95e655db2faf49d48f3055dc29eef189,
title = "Analysis of oligomeric protein complexes in the chloroplast sub-proteome of nucleic acid-binding proteins from mustard reveals potential redox regulators of plastid gene expression",
abstract = "Photosynthetic light quality acclimation in plants involves redox-controlled changes in plastid gene expression. To study proteins potentially involved in this regulation, we isolated low-abundant chloroplast nucleic acid-binding proteins from the crucifere mustard (Sinapis alba) and investigated if photosynthetic redox signals affect their composition and/or oligomeric structure. We purified chloroplasts from plants subjected to light quality shifts and applied organelle lysates to heparin-Sepharose chromatography followed by 2-D blue native PAGE. We studied accumulation and structure of oligomeric protein complexes and applied MS/MS to identify them. We found ten oligomeric protein complexes of higher order and eleven smaller protein complexes or spots including plastid-encoded RNA polymerase (PEP), plastid transcriptionally active chromosome proteins, RNA-binding proteins, ribosomal subunits and chaperones. A translation elongation factor was found to be the only protein displaying major differences in its amounts in response to the growth lights. Furthermore, we found a novel thioredoxin as a subunit of the PEP, a 2-Cys-peroxiredoxin complex and a (soluble) ferredoxin:NADP-oxido-reductase, which represent potential redox regulator of plastid gene expression. A T-DNA knock-out line of the thioredoxin from Arabidopsis exhibits a yellowish-pale phenotype, demonstrating that this novel PEP subunit is essential for proper plastid development.",
keywords = "Blotting, Western, DNA-Binding Proteins/metabolism, Mustard Plant/metabolism, Oxidation-Reduction, Plant Proteins/metabolism, Plastids/metabolism, Tandem Mass Spectrometry",
author = "Yvonne Schr{\"o}ter and Sebastian Steiner and Kevin Matth{\"a}i and Thomas Pfannschmidt",
year = "2010",
month = jun,
doi = "10.1002/pmic.200900678",
language = "English",
volume = "10",
pages = "2191--204",
journal = "PROTEOMICS",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "11",
}