title: Proteomics analysis reveals a highly heterogeneous proteasome composition and the post-translational regulation of peptidase activity under pathogen signaling in plants.
abstract: The proteasome is a large multisubunit complex that plays a crucial role in the removal of damaged or selective ubiquitinated proteins, thereby allowing quality control of cellular proteins and restricted regulation of diverse cellular signaling in eukaryotic cells. Proteasome-dependent protein degradation is involved in almost all aspects of plant growth and responses to environmental stresses including pathogen resistance. Although the molecular mechanism for specifying targets by ubiquitin ligases is well understood, the detailed characterization of the plant proteasome complex remains unclear. One of the most important features of the plant proteasome is that most subunits are encoded by duplicate genes, suggesting the highly heterogeneous composition of this proteasome. Here, we performed affinity purification and a combination of 2-dimensional electrophoresis and mass spectrometry, which identified the detailed composition of paralogous and modified proteins. Moreover, these proteomics approaches revealed that specific subunit composition and proteasome peptidase activity were affected by pathogen-derived MAMPs, flg22 treatment. Interestingly, flg22 treatment did not alter mRNA expression levels of the peptidase genes PBA, PBB1/2, PBE1/2, and total proteasome levels remained unchanged by flg22 as well. These results demonstrate the finely tuned mechanism that regulates proteasome function via putative post-translational modifications in response to environmental stress in plants.
authors.affiliation: Faculty of Science and Graduate School of Life Science, Hokkaido University , Sapporo 060-0810, Japan.
authors.forename: Hui H
authors.initials: HH
authors.lastname: Sun
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Fukao
authors.forename: Sakiko
authors.initials: S
authors.lastname: Ishida
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Shugo
authors.initials: S
authors.lastname: Maekawa
authors.forename: Masayuki
authors.initials: M
authors.lastname: Fujiwara
authors.forename: Takeo
authors.initials: T
authors.lastname: Sato
authors.forename: Junji
authors.initials: J
authors.lastname: Yamaguchi
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D002853:Chromatography, Liquid; D017931:DNA Primers / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D015151:Immunoblotting; D010447:Peptide Hydrolases / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D017931:DNA Primers; D010940:Plant Proteins; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex