, Volume 233, Issue 6, pp 1287–1292 | Cite as

Endogenous protein mono-ADP-ribosylation in Arabidopsis thaliana

  • Hai Wang
  • Qin Liang
  • Kaiming Cao
  • Xiaochun Ge
Rapid Communication


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.


Arabidopsis thaliana Mono-ADP-ribosylation Mono-ADP-ribosyltransferase Oxidative stress Post-translational modification 



Glyceraldehyde 3-phosphate dehydrogenase


Glutamate dehydrogenase




Poly (ADP-ribose) polymerase


Radical-induced cell death one 1


Similar to radical-induced cell death one 1



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.


  1. Ahlfors R, Lang S, Overmyer K, Jaspers P, Brosche M, Tauriainen A, Kollist H, Tuominen H, Belles-Boix E, Piippo M, Inze D, Palva ET, Kangasjarvi J (2004) Arabidopsis RADICAL-INDUCED CELL DEATH1 belongs to the WWE protein-protein interaction domain protein family and modulates abscisic acid, ethylene, and methyl jasmonate responses. Plant Cell 16(7):1925–1937PubMedCrossRefGoogle Scholar
  2. Aktories K (1997) Rho proteins: targets for bacterial toxins. Trends Microbiol 5(7):282–288PubMedCrossRefGoogle Scholar
  3. Ame JC, Spenlehauer C, de Murcia G (2004) The PARP superfamily. Bioessays 26(8):882–893PubMedCrossRefGoogle Scholar
  4. Cervantes-Laurean D, Loflin PT, Minter DE, Jacobson EL, Jacobson MK (1995) Protein modification by ADP-ribose via acid-labile linkages. J Biol Chem 270(14):7929–7936PubMedCrossRefGoogle Scholar
  5. Cervantes-Laurean D, Jacobson EL, Jacobson MK (1996) Glycation and glycoxidation of histones by ADP-ribose. J Biol Chem 271(18):10461–10469PubMedCrossRefGoogle Scholar
  6. Corda D, Di Girolamo M (2003) Functional aspects of protein mono-ADP-ribosylation. EMBO J 22(9):1953–1958PubMedCrossRefGoogle Scholar
  7. Dani N, Stilla A, Marchegiani A, Tamburro A, Till S, Ladurner AG, Corda D, Di Girolamo M (2009) Combining affinity purification by ADP-ribose-binding macro domains with mass spectrometry to define the mammalian ADP-ribosyl proteome. Proc Natl Acad Sci USA 106(11):4243–4248PubMedCrossRefGoogle Scholar
  8. De Block M, Verduyn C, De Brouwer D, Cornelissen M (2005) Poly(ADP-ribose) polymerase in plants affects energy homeostasis, cell death and stress tolerance. Plant J 41(1):95–106PubMedCrossRefGoogle Scholar
  9. Di Girolamo M, Dani N, Stilla A, Corda D (2005) Physiological relevance of the endogenous mono(ADP-ribosyl)ation of cellular proteins. FEBS J 272(18):4565–4575PubMedCrossRefGoogle Scholar
  10. Frei B, Richter C (1988) Mono(ADP-ribosylation) in rat liver mitochondria. Biochemistry 27(2):529–535PubMedCrossRefGoogle Scholar
  11. Fu ZQ, Guo M, Jeong BR, Tian F, Elthon TE, Cerny RL, Staiger D, Alfano JR (2007) A type III effector ADP-ribosylates RNA-binding proteins and quells plant immunity. Nature 447(7142):284–288PubMedCrossRefGoogle Scholar
  12. Fujibe T, Saji H, Arakawa K, Yabe N, Takeuchi Y, Yamamoto KT (2004) A methyl viologen-resistant mutant of Arabidopsis, which is allelic to ozone-sensitive rcd1, is tolerant to supplemental ultraviolet-B irradiation. Plant Physiol 134(1):275–285PubMedCrossRefGoogle Scholar
  13. Herrero-Yraola A, Bakhit SM, Franke P, Weise C, Schweiger M, Jorcke D, Ziegler M (2001) Regulation of glutamate dehydrogenase by reversible ADP-ribosylation in mitochondria. EMBO J 20(10):2404–2412PubMedCrossRefGoogle Scholar
  14. Jacobson EL, Cervantes-Laurean D, Jacobson MK (1994) Glycation of proteins by ADP-ribose. Mol Cell Biochem 138(1–2):207–212PubMedCrossRefGoogle Scholar
  15. Jacobson EL, Cervantes-Laurean D, Jacobson MK (1997) ADP-ribose in glycation and glycoxidation reactions. Adv Exp Med Biol 419:371–379PubMedGoogle Scholar
  16. Jaspers P, Blomster T, Brosche M, Salojarvi J, Ahlfors R, Vainonen JP, Reddy RA, Immink R, Angenent G, Turck F, Overmyer K, Kangasjarvi J (2009) Unequally redundant RCD1 and SRO1 mediate stress and developmental responses and interact with transcription factors. Plant J 60(2):268–279PubMedCrossRefGoogle Scholar
  17. Jaspers P, Overmyer K, Wrzaczek M, Vainonen JP, Blomster T, Salojarvi J, Reddy RA, Kangasjarvi J (2010) The RST and PARP-like domain containing SRO protein family: analysis of protein structure, function and conservation in land plants. BMC Genomics 11:170PubMedCrossRefGoogle Scholar
  18. Jorcke D, Ziegler M, Herrero-Yraola A, Schweiger M (1998) Enzymic, cysteine-specific ADP-ribosylation in bovine liver mitochondria. Biochem J 332(Pt 1):189–193PubMedGoogle Scholar
  19. Katiyar-Agarwal S, Zhu J, Kim K, Agarwal M, Fu X, Huang A, Zhu JK (2006) The plasma membrane Na +/H + antiporter SOS1 interacts with RCD1 and functions in oxidative stress tolerance in Arabidopsis. Proc Natl Acad Sci USA 103(49):18816–18821PubMedCrossRefGoogle Scholar
  20. Ledford BE, Leno GH (1994) ADP-ribosylation of the molecular chaperone GRP78/BiP. Mol Cell Biochem 138(1–2):141–148PubMedCrossRefGoogle Scholar
  21. Liu Y, Kahn ML (1995) ADP-ribosylation of Rhizobium meliloti glutamine synthetase III in vivo. J Biol Chem 270(4):1624–1628PubMedCrossRefGoogle Scholar
  22. Ludden PW (1994) Reversible ADP-ribosylation as a mechanism of enzyme regulation in procaryotes. Mol Cell Biochem 138(1–2):123–129PubMedCrossRefGoogle Scholar
  23. Lupi R, Corda D, Di Girolamo M (2000) Endogenous ADP-ribosylation of the G protein beta subunit prevents the inhibition of type 1 adenylyl cyclase. J Biol Chem 275(13):9418–9424PubMedCrossRefGoogle Scholar
  24. McDonald LJ, Moss J (1994) Enzymatic and nonenzymatic ADP-ribosylation of cysteine. Mol Cell Biochem 138(1–2):221–226PubMedCrossRefGoogle Scholar
  25. Mekalanos JJ, Collier RJ, Romig WR (1979) Enzymic activity of cholera toxin. I. New method of assay and the mechanism of ADP-ribosyl transfer. J Biol Chem 254(13):5849–5854PubMedGoogle Scholar
  26. Mendoza-Alvarez H, Alvarez-Gonzalez R (1993) Poly(ADP-ribose) polymerase is a catalytic dimer and the automodification reaction is intermolecular. J Biol Chem 268(30):22575–22580PubMedGoogle Scholar
  27. Moss J, Stanley SJ, Nightingale MS, Murtagh JJ Jr, Monaco L, Mishima K, Chen HC, Williamson KC, Tsai SC (1992) Molecular and immunological characterization of ADP-ribosylarginine hydrolases. J Biol Chem 267(15):10481–10488PubMedGoogle Scholar
  28. Overmyer K, Tuominen H, Kettunen R, Betz C, Langebartels C, Sandermann H Jr, Kangasjarvi J (2000) Ozone-sensitive arabidopsis rcd1 mutant reveals opposite roles for ethylene and jasmonate signaling pathways in regulating superoxide-dependent cell death. Plant Cell 12(10):1849–1862PubMedCrossRefGoogle Scholar
  29. Saxty BA, van Heyningen S (1995) The purification of a cysteine-dependent NAD + glycohydrolase activity from bovine erythrocytes and evidence that it exhibits a novel ADP-ribosyltransferase activity. Biochem J 310(Pt 3):931–937PubMedGoogle Scholar
  30. Tanuma S, Kawashima K, Endo H (1988) Eukaryotic mono(ADP-ribosyl)transferase that ADP-ribosylates GTP-binding regulatory Gi protein. J Biol Chem 263(11):5485–5489PubMedGoogle Scholar
  31. Terashima M, Yamamori C, Shimoyama M (1995) ADP-ribosylation of Arg28 and Arg206 on the actin molecule by chicken arginine-specific ADP-ribosyltransferase. Eur J Biochem 231(1):242–249PubMedCrossRefGoogle Scholar
  32. Till S, Diamantara K, Ladurner AG (2008) PARP: a transferase by any other name. Nat Struct Mol Biol 15(12):1243–1244PubMedCrossRefGoogle Scholar
  33. Van Ness BG, Barrowclough B, Bodley JW (1980) Recognition of elongation factor 2 by diphtheria toxin is not solely defined by the presence of diphthamide. FEBS Lett 120(1):4–6PubMedCrossRefGoogle Scholar
  34. Villamon E, Gozalbo D, Martinez JP, Gil ML (1999) Purification of a biologically active recombinant glyceraldehyde 3-phosphate dehydrogenase from Candida albicans. FEMS Microbiol Lett 179(1):61–65PubMedCrossRefGoogle Scholar
  35. Yamaizumi M, Uchida T, Okada Y, Furusawa M (1978) Neutralization of diphtheria toxin in living cells by microinjection of antifragment A contained within resealed erythrocyte ghosts. Cell 13(2):227–232PubMedCrossRefGoogle Scholar
  36. Zolkiewska A, Moss J (1993) Integrin alpha 7 as substrate for a glycosylphosphatidylinositol-anchored ADP-ribosyltransferase on the surface of skeletal muscle cells. J Biol Chem 268(34):25273–25276PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.State Key Laboratory of Genetic Engineering, Department of Biochemistry, Institute of Plant Biology, School of Life SciencesFudan UniversityShanghaiChina

Personalised recommendations