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Endogenous protein mono-ADP-ribosylation in Arabidopsis thaliana

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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.

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Abbreviations

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

GDH:

Glutamate dehydrogenase

mART:

Mono-ADP-ribosyltransferase

PARP:

Poly (ADP-ribose) polymerase

RCD1:

Radical-induced cell death one 1

SRO1:

Similar to radical-induced cell death one 1

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 30670178, 30770425 and 31070232) and Wang Dao Scholar Plan sponsored by Fudan University. We thank Professor Hong Ma at Fudan University for helpful suggestions and manuscript editing.

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Correspondence to Xiaochun Ge.

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Wang, H., Liang, Q., Cao, K. et al. Endogenous protein mono-ADP-ribosylation in Arabidopsis thaliana . Planta 233, 1287–1292 (2011). https://doi.org/10.1007/s00425-011-1415-y

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