title: Endogenous protein mono-ADP-ribosylation in Arabidopsis thaliana.
abstract: Protein mono-ADP-ribosylation post-translationally transfers the ADP-ribose moiety from the β-NAD+ donor to various protein acceptors. This type of modification has been widely characterized and shown to regulate protein activities in animals, yeast and prokaryotes, but has never been reported in plants. In this study, using [³²P]NAD+ as the substrate, ADP-ribosylated proteins in Arabidopsis were investigated. One protein substrate of 32 kDa in adult rosette leaves was found to be radiolabeled. Heat treatment, protease sensitivity and nucleotide derivative competition assays suggested a covalent reaction of NAD+ with the 32 kDa protein. [carbonyl-¹⁴C]NAD+ could not label the 32 kDa protein, confirming that the modification was ADP-ribosylation. Poly (ADP-ribose) polymerase inhibitor failed to suppress the reaction, but chemicals that destroy mono-ADP-ribosylation on specific amino acid residues could break up the linkage, suggesting that the reaction was not a poly-ADP-ribosylation but rather a mono-ADP-ribosylation. This modification mainly existed in leaves and was enhanced by oxidative stresses. In young seedlings, two more protein substrates with the size of 45 kDa and over 130 kDa, respectively, were observed in addition to the 32 kDa protein, indicating that different proteins were modified at different developmental stages. Although the substrate proteins remain to be identified, this is the first report on the characterization of endogenously mono-ADP-ribosylated proteins in plants.
authors.affiliation: State Key Laboratory of Genetic Engineering, Department of Biochemistry, Institute of Plant Biology, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, China.
authors.forename: Hai
authors.initials: H
authors.lastname: Wang
authors.forename: Qin
authors.initials: Q
authors.lastname: Liang
authors.forename: Kaiming
authors.initials: K
authors.lastname: Cao
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Ge
mesh_terms: D036002:ADP Ribose Transferases / Q000378:metabolism*; D020727:ADP-Ribosylation Factors; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006031:Glycosylation; D009243:NAD / Q000378:metabolism; D009928:Organ Specificity; D018384:Oxidative Stress / Q000502:physiology; D018515:Plant Leaves / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000502:physiology
chemical_list: D009243:NAD; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; D020727:ADP-Ribosylation Factors; D003545:Cysteine