Protein & Cell

, Volume 3, Issue 6, pp 450–459 | Cite as

The enzymatic activity of Arabidopsis protein arginine methyltransferase 10 is essential for flowering time regulation

  • Lifang Niu
  • Falong Lu
  • Taolan Zhao
  • Chunyan Liu
  • Xiaofeng Cao
Research Article

Abstract

Arabidopsis AtPRMT10 is a plant-specific type I protein arginine methyltransferase that can asymmetrically dimethylate arginine 3 of histone H4 with auto-methylation activity. Mutations of AtPRMT10 derepress FLOWERINGLOCUS C (FLC) expression resulting in a late-flowering phenotype. Here, to further investigate the biochemical characteristics of AtPRMT10, we analyzed a series of mutated forms of the AtPRMT10 protein. We demonstrate that the conserved “VLD” residues and “double-E loop” are essential for enzymatic activity of AtPRMT10. In addition, we show that Arg54 and Cys259 of AtPRMT10, two residues unreported in animals, are also important for its enzymatic activity. We find that Arg13 of AtPRMT10 is the auto-methylation site. However, substitution of Arg13 to Lys13 does not affect its enzymatic activity. In vivo complementation assays reveal that plants expressing AtPRMT10 with VLD-AAA, E143Q or E152Q mutations retain high levels of FLC expression and fail to rescue the late-flowering phenotype of atprmt10 plants. Taken together, we conclude that the methyltransferase activity of AtPRMT10 is essential for repressing FLC expression and promoting flowering in Arabidopsis.

Keywords

protein arginine methyltransferases (PRMTs) flowering methyltransferase activity 

Supplementary material

13238_2012_2935_MOESM1_ESM.pdf (865 kb)
Supplementry Material(PDF 865 kb)

References

  1. Ahmad, A., and Cao, X. (2012). Plant PRMTs broaden the scope of arginine methylation. J Genet Genomics (In Press).Google Scholar
  2. Ahmad, A., Zhang, Y., and Cao, X.F. (2010). Decoding the epigenetic language of plant development. Mol Plant 3, 719–728.CrossRefGoogle Scholar
  3. Amasino, R.M. (2005). Vernalization and flowering time. Curr Opin Biotechnol 16, 154–158.CrossRefGoogle Scholar
  4. Ausin, I., Alonso-Blanco, C., Jarillo, J.A., Ruiz-Garcia, L., and Martinez-Zapater, J.M. (2004). Regulation of flowering time by FVE, a retinoblastoma-associated protein. Nat Genet 36, 162–166.CrossRefGoogle Scholar
  5. Bechtold, N., and Pelletier, G. (1998). In planta agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Arabidopsis Protoc 82, 259–266.CrossRefGoogle Scholar
  6. Bedford, M.T., and Clarke, S.G. (2009). Protein arginine methylation in mammals: who, what, and why. Mol Cell 33, 1–13.CrossRefGoogle Scholar
  7. Cheng, X., Collins, R., and Zhang, X. (2005). Structural and sequence motifs of protein (histone) methylation enzymes. Annu Rev Biophys Biomol Struct 34, 267–294.CrossRefGoogle Scholar
  8. Cheng, Y., Frazier, M., Lu, F., Cao, X., and Redinbo, M.R. (2011). Crystal structure of the plant epigenetic protein arginine methyltransferase 10. J Mol Biol 414, 106–122.CrossRefGoogle Scholar
  9. Deng, X., Gu, L., Liu, C., Lu, T., Lu, F., Lu, Z., Cui, P., Pei, Y., Wang, B., Hu, S., et al. (2010). Arginine methylation mediated by the Arabidopsis homolog of PRMT5 is essential for proper pre-mRNA splicing. Proc Natl Acad Sci U S A 107, 19114–19119.CrossRefGoogle Scholar
  10. Frankel, A., Yadav, N., Lee, J., Branscombe, T.L., Clarke, S., and Bedford, M.T. (2002). The novel human protein arginine N-methyltransferase PRMT6 is a nuclear enzyme displaying unique substrate specificity. J Biol Chem 277, 3537–3543.CrossRefGoogle Scholar
  11. Gui, S., Wooderchak, W.L., Daly, M.P., Porter, P.J., Johnson, S.J., and Hevel, J.M. (2011). Investigation of the molecular origins of protein-arginine methyltransferase I (PRMT1) product specificity reveals a role for two conserved methionine residues. J Biol Chem 286, 29118–29126.CrossRefGoogle Scholar
  12. He, Y. (2009). Control of the transition to flowering by chromatin modifications. Mol Plant 2, 554–564.CrossRefGoogle Scholar
  13. Herrmann, F., and Fackelmayer, F. (2009). Nucleo-cytoplasmic shuttling of protein arginine methyltransferase 1 (PRMT1) requires enzymatic activity. Genes Cells 14, 309–317.CrossRefGoogle Scholar
  14. Hess, D.T., Matsumoto, A., Kim, S.O., Marshall, H.E., and Stamler, J.S. (2005). Protein S-nitrosylation: purview and parameters. N at Rev Mol Cell Biol 6, 150–166.CrossRefGoogle Scholar
  15. Higashimoto, K., Kuhn, P., Desai, D., Cheng, X., and Xu, W. (2007). Phosphorylation-mediated inactivation of coactivator-associated arginine methyltransferase 1. Proc Natl Acad Sci U S A 104, 12318–12323.CrossRefGoogle Scholar
  16. Hong, S., Song, H.R., Lutz, K., Kerstetter, R.A., Michael, T.P., and McClung, C.R. (2010). Type II protein arginine methyltransferase 5 (PRMT5) is required for circadian period determination in Arabidopsis thaliana. Proc Natl Acad Sci U S A 107, 21211–21216.CrossRefGoogle Scholar
  17. Hwang, H., Pierce, B., Mintseris, J., Janin, J., and Weng, Z. (2008). Protein-protein docking benchmark version 3.0. Proteins 73, 705–709.CrossRefGoogle Scholar
  18. Jiang, D., Yang, W., He, Y., and Amasino, R.M. (2007). Arabidopsis relatives of the human lysine-specific demethylase1 repress the expression of FWA and FLOWERING LOCUS C and thus promote the floral transition. Plant Cell 19, 2975–2987.CrossRefGoogle Scholar
  19. Kuhn, P., Chumanov, R., Wang, Y., Ge, Y., Burgess, R.R., and Xu, W. (2011). Automethylation of CARM1 allows coupling of transcription and mRNA splicing. Nucleic Acids Res 39, 2717–2726.CrossRefGoogle Scholar
  20. Kuhn, P., Xu, Q., Cline, E., Zhang, D., Ge, Y., and Xu, W. (2009). Delineating Anopheles gambiae coactivator associated arginine methyltransferase 1 automethylation using top-down high resolution tandem mass spectrometry. Protein Sci 18, 1272–1280.CrossRefGoogle Scholar
  21. Kwak, Y.T., Guo, J., Prajapati, S., Park, K.J., Surabhi, R.M., Miller, B., Gehrig, P., and Gaynor, R.B. (2003). Methylation of SPT5 regulates its interaction with RNA polymerase II and transcriptional elongation properties. Mol Cell 11, 1055–1066.CrossRefGoogle Scholar
  22. Lee, I., Aukerman, M., Gore, S., Lohman, K., Michaels, S., Weaver, L., John, M., Feldmann, K., and Amasino, R. (1994). Isolation of LUMINIDEPENDENS: a gene involved in the control of flowering time in Arabidopsis. The Plant Cell 6, 75–83.CrossRefGoogle Scholar
  23. Lee, Y.H., Koh, S.S., Zhang, X., Cheng, X., and Stallcup, M.R. (2002). Synergy among nuclear receptor coactivators: selective requirement for protein methyltransferase and acetyltransferase activities. Mol Cell Biol 22, 3621–3632.CrossRefGoogle Scholar
  24. Li, F., Huarte, M., Zaratiegui, M., Vaughn, M.W., Shi, Y., Martienssen, R., and Cande, W.Z. (2008). Lid2 is required for coordinating H3K4 and H3K9 methylation of heterochromatin and euchromatin. Cell 135, 272–283.CrossRefGoogle Scholar
  25. Lim, M.H., Kim, J., Kim, Y.S., Chung, K.S., Seo, Y.H., Lee, I., Hong, C.B., Kim, H.J., and Park, C.M. (2004). A new Arabidopsis gene, FLK, encodes an RNA binding protein with K homology motifs and regulates flowering time via FLOWERING LOCUS C. Plant Cell 16, 731–740.CrossRefGoogle Scholar
  26. Liu, C., Lu, F., Cui, X., and Cao, X. (2010). Histone methylation in higher plants. Annu Rev Plant Biol 61, 395–420.CrossRefGoogle Scholar
  27. Macknight, R., Bancroft, I., Page, T., Lister, C., Schmidt, R., Love, K., Westphal, L., Murphy, G., Sherson, S., and Cobbett, C. (1997). FCA, a gene controlling flowering time in Arabidopsis, encodes a protein containing RNA-binding domains. Cell 89, 737–745.CrossRefGoogle Scholar
  28. Michaels, S.D., and Amasino, R.M. (1999). FLOWERING LOCUS C encodes a novel MADS domain protein that acts as a repressor of flowering. Plant Cell 11, 949–956.CrossRefGoogle Scholar
  29. Niu, L., Lu, F., Pei, Y., Liu, C., and Cao, X. (2007). Regulation of flowering time by the protein arginine methyltransferase AtPRMT10. EMBO Rep 8, 1190–1195.CrossRefGoogle Scholar
  30. Niu, L., Zhang, Y., Pei, Y., Liu, C., and Cao, X. (2008). Redundant requirement for a pair of PROTEIN ARGININE METHYLTRANSFERASE4 homologs for the proper regulation of Arabidopsis flowering time. Plant Physiol 148, 490–503.CrossRefGoogle Scholar
  31. Noh, B., Lee, S.H., Kim, H.J., Yi, G., Shin, E.A., Lee, M., Jung, K.J., Doyle, M.R., Amasino, R.M., and Noh, Y.S. (2004). Divergent roles of a pair of homologous jumonji/zinc-finger-class transcription factor proteins in the regulation of Arabidopsis flowering time. Plant Cell 16, 2601–2613.CrossRefGoogle Scholar
  32. Pei, Y., Niu, L., Lu, F., Liu, C., Zhai, J., Kong, X., and Cao, X. (2007). Mutations in the Type II protein arginine methyltransferase AtPRMT5 result in pleiotropic developmental defects in Arabidopsis. Plant Physiol 144, 1913–1923.CrossRefGoogle Scholar
  33. Quesada, V., Dean, C., and Simpson, G.G. (2005). Regulated RNA processing in the control of Arabidopsis flowering. Int J Dev Biol 49, 773–780.CrossRefGoogle Scholar
  34. Sanchez, S.E., Petrillo, E., Beckwith, E.J., Zhang, X., Rugnone, M.L., Hernando, C.E., Cuevas, J.C., Godoy Herz, M.A., Depetris-Chauvin, A., Simpson, C.G., et al. (2010). A methyl transferase links the circadian clock to the regulation of alternative splicing. Nature 468, 112–116.CrossRefGoogle Scholar
  35. Sayegh, J., Webb, K., Cheng, D., Bedford, M.T., and Clarke, S.G. (2007). Regulation of protein arginine methyltransferase 8 (PRMT8) activity by its N-terminal domain. J Biol Chem 282, 36444–36453.CrossRefGoogle Scholar
  36. Schmitz, R.J., Sung, S., and Amasino, R.M. (2008). Histone arginine methylation is required for vernalization-induced epigenetic silencing of FLC in winter-annual Arabidopsis thaliana. Proc Natl Acad Sci U S A 105, 411–416.CrossRefGoogle Scholar
  37. Schluckebier, G., O’Gara, M., Saenger, W., and Cheng, X. (1995). Universal catalytic domain structure of AdoMet-dependent methyltransferases. J Mol Biol 247, 16–20.CrossRefGoogle Scholar
  38. Schomburg, F.M., Patton, D.A., Meinke, D.W., and Amasino, R.M. (2001). FPA, a gene involved in floral induction in Arabidopsis, encodes a protein containing RNA-recognition motifs. Plant Cell 13, 1427–1436.CrossRefGoogle Scholar
  39. Simpson, G.G. (2004). The autonomous pathway: epigenetic and post-transcriptional gene regulation in the control of Arabidopsis flowering time. Curr Opin Plant Biol 7, 570–574.CrossRefGoogle Scholar
  40. Simpson, G.G., Dijkwel, P.P., Quesada, V., Henderson, I., and Dean, C. (2003). FY is an RNA 3′ end-processing factor that interacts with FCA to control the Arabidopsis floral transition. Cell 1113, 777–787.CrossRefGoogle Scholar
  41. Tada, Y., Spoel, S.H., Pajerowska-Mukhtar, K., Mou, Z., Song, J., Wang, C., Zuo, J., and Dong, X. (2008). Plant immunity requires conformational charges of NPR1 via S-nitrosylation and thioredoxins. Science 321, 952–956.CrossRefGoogle Scholar
  42. Wada, K., Inoue, K., and Hagiwara, M. (2002). Identification of methylated proteins by protein arginine N-methyltransferase 1, PRMT1, with a new expression cloning strategy. Biochim Biophys Acta 1591, 1–10.CrossRefGoogle Scholar
  43. Wang, X., Zhang, Y., Ma, Q., Zhang, Z., Xue, Y., Bao, S., and Chong, K. (2007). SKB1-mediated symmetric dimethylation of histone H4R3 controls flowering time in Arabidopsis. EMBO J 26, 1934–1941.CrossRefGoogle Scholar
  44. Weiss, V., McBride, A., Soriano, M., Filman, D., Silver, P., and Hogle, J. (2000). The structure and oligomerization of the yeast arginine methyltransferase, Hmt1. Nat Struct Biol 7, 1165–1171.CrossRefGoogle Scholar
  45. Wolf, S.S. (2009). The protein arginine methyltransferase family: an update about function, new perspectives and the physiological role in humans. Cell Mol Life Sci 66, 2109–2121.CrossRefGoogle Scholar
  46. Xu, W., Cho, H., Kadam, S., Banayo, E.M., Anderson, S., Yates, J.R., Emerson, B.M., and Evans, R.M. (2004). A methylation-mediator complex in hormone signaling. Genes Dev 18, 144–156.CrossRefGoogle Scholar
  47. Yan, D., Zhang, Y., Niu, L., Yuan, Y., and Cao, X. (2007). Identification and characterization of two closely related histone H4 arginine 3 methyltransferases in Arabidopsis thaliana. Biochem J 408, 113–121.CrossRefGoogle Scholar
  48. Zhang, J., Teng, C., and Liang, Y. (2011a). Programmed cell death may act as a surveillance mechanism to safeguard male gametophyte development in Arabidopsis. Protein Cell 2, 837–844.CrossRefGoogle Scholar
  49. Zhang, X., and Cheng, X. (2003). Structure of the predominant protein arginine methyltransferase PRMT1 and analysis of its binding to substrate peptides. Structure 11, 509–520.CrossRefGoogle Scholar
  50. Zhang, X., Zhou, L., and Cheng, X. (2000). Crystal structure of the conserved core of protein arginine methyltransferase PRMT3. EMBO J 19, 3509–3519.CrossRefGoogle Scholar
  51. Zhang, Z., Zhang, S., Zhang, Y., Wang, X., Li, D., Li, Q., Yue, M., Zhang, Y.E., Xu, Y., Xue, Y., et al. (2011b). Arabidopsis floral initiator SKB1 confers high salt tolerance by regulating transcription and pre-mRNA splicing through altering histone H4R3 and small nuclear ribonucleoprotein LSM4 methylation. Plant Cell 23, 396–411.CrossRefGoogle Scholar

Copyright information

© Higher Education Press and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Lifang Niu
    • 1
    • 2
    • 4
  • Falong Lu
    • 1
    • 3
    • 4
  • Taolan Zhao
    • 1
    • 4
  • Chunyan Liu
    • 1
  • Xiaofeng Cao
    • 1
  1. 1.State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijingChina
  2. 2.Institute for Agricultural BiosciencesOklahoma State UniversityArdmoreUSA
  3. 3.Lineberger Comprehensive Cancer CenterUniversity of North Carolina at Chapel HillChapel HillUSA
  4. 4.Graduate School of the Chinese Academy of SciencesBeijingChina

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