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Plant Molecular Biology

, Volume 80, Issue 4–5, pp 461–476 | Cite as

Novel role for a serine/arginine-rich splicing factor, AdRSZ21 in plant defense and HR-like cell death

  • Koppolu Raja Rajesh Kumar
  • P. B. Kirti
Article

Abstract

A splicing factor gene belonging to the serine/arginine (SR)-rich protein family was cloned from Arachis diogoi, a wild relative of peanut in a study on differential gene expression and was designated as AdRSZ21. AdRSZ21 exhibits a RNA recognition motif (RRM), a CCHC type zinc finger domain (Zinc Knuckle, ZnK) and a C-terminal RS domain that is rich in arginine and serine. Multiple sequence alignment of AdRSZ21 with putative orthologs from diverse taxa including lower plants and monocots showed that the RRM and ZnK domains are evolutionarily conserved. Phylogenetic studies revealed that AdRSZ21 belongs to the RSZ subfamily and is closely related to the Arabidopsis ortholog AtRSZ22. Transient constitutive and conditional heterologous expression of AdRSZ21 resulted in HR-like cell death in tobacco leaves. The presence of a functional RRM domain, but not ZnK domain was essential for AdRSZ21 induced HR-like cell death phenotype. On the other hand, expression of AdRSZ21 with mutated ZnK domain lead to accelerated cell death. The cell death induced by AdRSZ21 was found to be associated with specific upregulation of patatin-like protein gene and other defense related gene transcripts suggesting a role for AdRSZ21 in plant defense and HR-like cell death.

Keywords

Serine/arginine-rich protein Alternative splicing Arachis diogoi Programmed cell death Patatin-like protein Ribosomal protein Phaeoisariopsis personata 

Notes

Acknowledgments

The work was supported by a grant (BT/PR6853/PBD/16/627/2005) from the Department of Biotechnology, Government of India. Arachis wild germplasm used in this study has been kindly provided by ICRISAT, Patancheru, India. The authors thank Prof. Nam-Hai Chua, Rockefeller University, New York, USA for pER8 vector. The authors would like to thank Dr. K. Uma Mahendra Kumar, IGCAR, Kalpakkam for helping with the analysis of IDRs. The authors also acknowledge the research facilities supported by DST-FIST, UGC-CAS etc. to the Department of Plant Sciences, University of Hyderabad. KRRK was supported by Dr. D.S.Kothari Postdoctoral Fellowship from University Grants Commission (UGC), Government of India.

Supplementary material

11103_2012_9960_MOESM1_ESM.pdf (1.1 mb)
Supplementary material 1 (PDF 1084 kb)

References

  1. Ali GS, Palusa SG, Golovkin M, Prasad J, Manley JL et al (2007) Regulation of plant developmental processes by a novel splicing factor. PLoS ONE 2(5):e471. doi: 10.1371/journal.pone.0000471 PubMedCrossRefGoogle Scholar
  2. Baker CJ, Mock NM (1994) An improved method for monitoring cell death in cell suspension and leaf disc assays using Evans blue. Plant Cell Tissue Organ Cult 39:7–12CrossRefGoogle Scholar
  3. Barta A, Kalyna M, Reddy ASN (2010) Implementing a rational and consistent nomenclature for serine/arginine-rich protein splicing factors (SR proteins) in plants. Plant Cell 22:2926–2929PubMedCrossRefGoogle Scholar
  4. Brown CJ, Takayama S, Campen AM, Vise P, Marshall TW, Oldfield CJ, Williams CJ, Keith Dunker A (2002) Evolutionary rate heterogeneity in proteins with long disordered regions. J Mol Evol 55:104–110Google Scholar
  5. Caceres JF, Krainer AR (1993) Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains. EMBO J 12:4715–4726PubMedGoogle Scholar
  6. Cáceres JF, Screaton GR, Krainer AR (1998) A specific subset of SR proteins shuttles continuously between the nucleus and the cytoplasm. Genes Dev 12:55–66PubMedCrossRefGoogle Scholar
  7. Canonne J, Froidure-Nicolas S, Rivas S (2011) Phospholipases in action during plant defense signaling. Plant Signal Behav 6:13–18PubMedCrossRefGoogle Scholar
  8. Carvalho RF, Carvalho SD, Duque P (2010) The Plant-Specific SR45 protein negatively regulates glucose and ABA signaling during early seedling development in Arabidopsis. Plant Physiol 154:772–783PubMedCrossRefGoogle Scholar
  9. Cavaloc Y, Bourgeois CF, Kiste L, Stévenin J (1999) The splicing factors 9G8 and SRp20 transactivate splicing through different and specific enhancers. RNA 5:468–483PubMedCrossRefGoogle Scholar
  10. Coll NS, Epple P, Dangl JL (2011) Programmed cell death in the plant immune system. Cell Death Differ 18:1247–1256PubMedCrossRefGoogle Scholar
  11. Degenhardt RF, Bonham-Smith PC (2008) Arabidopsis ribosomal proteins RPL23aA and RPL23aB are differentially targeted to the nucleolus and are disparately required for normal development. Plant Physiol 147:128–142PubMedCrossRefGoogle Scholar
  12. Dhondt S, Geoffroy P, Stelmach BA, Legrand M, Heitz T (2000) Soluble phospholipase A2 activity is induced before oxylipin accumulation in tobacco mosaic virus-infected tobacco leaves and is contributed by patatin-like enzymes. Plant J 23:431–440PubMedCrossRefGoogle Scholar
  13. Dhondt S, Gouzerh G, Müller A, Legrand M, Heitz T (2002) Spatio-temporal expression of patatin-like lipid acyl hydrolases and accumulation of jasmonates in elicitor-treated tobacco leaves are not affected by endogenous levels of salicylic acid. Plant J 32:749–762PubMedCrossRefGoogle Scholar
  14. Dinesh-Kumar SP, Baker BJ (2000) Alternatively spliced N resistance gene transcripts: their possible role in tobacco mosaic virus resistance. Proc Natl Acad Sci USA 97:1908–1913PubMedCrossRefGoogle Scholar
  15. Ding S, Shi J, Qian W, Iqbal K, Grundke-Iqbal I, Gong C-X, Liu F (2012) Regulation of alternative splicing of tau exon 10 by 9G8 and Dyrk1A. Neurobiol Aging 33:1389–1399PubMedCrossRefGoogle Scholar
  16. Dosztányi Z, Csizmok V, Tompa P, Simon I (2005) IUPred: Web server for the prediction of intrinsically unstructured regions of proteins based on estimated energy content. Bioinformatics 21:3433–3434PubMedCrossRefGoogle Scholar
  17. Duque P (2011) A role for SR proteins in plant stress responses. Plant Signal Behav 6:49–54PubMedCrossRefGoogle Scholar
  18. Feilner T, Hultschig C, Lee J, Meyer S, Immink RGH, Koenig A et al (2005) High throughput identification of potential Arabidopsis mitogen-activated protein kinases substrates. Mol Cell Proteomics 4:1558–1568PubMedCrossRefGoogle Scholar
  19. Filichkin SA, Priest HD, Givan SA, Shen R, Bryant DW, Fox SE et al (2010) Genome-wide mapping of alternative splicing in Arabidopsis thaliana. Genome Res 20:45–58PubMedCrossRefGoogle Scholar
  20. Garcia-Blanco MA, Baraniak AP, Lasda EL (2004) Alternative splicing in disease and therapy. Nat Biotechnol 22:535–546PubMedCrossRefGoogle Scholar
  21. Gilbert W, Siebel CW, Guthrie C (2001) Phosphorylation by Sky1p promotes Npl3p shuttling and mRNA dissociation. RNA M7:302–313CrossRefGoogle Scholar
  22. Hargous Y, Hautbergue GM, Tintaru AM, Skrisovska L, Golovanov AP, Stevenin J et al (2006) Molecular basis of RNA recognition and TAP binding by the SR proteins SRp20 and 9G8. EMBO J 25:5126–5137PubMedCrossRefGoogle Scholar
  23. Isshiki M, Tsumoto A, Shimamoto K (2006) The serine/arginine-rich protein family in rice plays important roles in constitutive and alternative splicing of pre-mRNA. Plant Cell 18:146–158PubMedCrossRefGoogle Scholar
  24. Kalyna M, Lopato S, Barta A (2003) Ectopic expression of atRSZ33 reveals its function in splicing and causes pleiotropic changes in development. Mol Biol Cell 14:3565–3577PubMedCrossRefGoogle Scholar
  25. Karni R, De Stanchina E, Lowe SW, Sinha R, Mu D, Krainer AR (2007) The gene encoding the splicing factor SF2/ASF is a proto-oncogene. Nat Struct Mol Biol 14:185–193PubMedCrossRefGoogle Scholar
  26. Karrer EE, Beachy RN, Holt CA (1998) Cloning of tobacco genes that elicit the hypersensitive response. Plant Mol Biol 36:681–690PubMedCrossRefGoogle Scholar
  27. Kumar KRR, Kirti PB (2010) A mitogen-activated protein kinase, AhMPK6 from peanut localizes to the nucleus and also induces defense responses upon transient expression in tobacco. Plant Physiol Biochem 48:481–486PubMedCrossRefGoogle Scholar
  28. Kumar KRR, Kirti PB (2011) Differential gene expression in Arachis diogoi upon interaction with peanut late leaf spot pathogen, Phaeoisariopsis personata and characterization of a pathogen induced cyclophilin. Plant Mol Biol 75:497–513PubMedCrossRefGoogle Scholar
  29. La Camera S, Balagué C, Göbel C, Geoffroy P, Legrand M et al (2009) The Arabidopsis patatin-like protein 2 (PLP2) plays an essential role in cell death execution and differentially affects biosynthesis of oxylipins and resistance to pathogens. Mol Plant Microbe Interact 22:469–481Google Scholar
  30. Lazar G, Schaal T, Maniatis T, Goodman HM (1995) Identification of a plant serine–arginine-rich protein similar to the mammalian splicing factor SF2/ASF. Proc Natl Acad Sci USA 92:7672–7676PubMedCrossRefGoogle Scholar
  31. Letunic I, Bork P (2011) Interactive tree of life v2: online annotation and display of phylogenetic trees made easy. Nucleic Acids Res 39:W475–W478Google Scholar
  32. Li J, Brader G, Palva ET (2008) Kunitz trypsin inhibitor: an antagonist of cell death triggered by phytopathogens and fumonisin B1 in Arabidopsis. Mol Plant 1:482–495PubMedCrossRefGoogle Scholar
  33. Liu Y, Zhang S (2004) Phosphorylation of 1-aminocyclopropane-1-carboxylic acid synthase by MPK6, a stress-responsive mitogen-activated protein kinase, induces ethylene biosynthesis in arabidopsis. Plant Cell 16:3386–3399PubMedCrossRefGoogle Scholar
  34. Liu H, Wang Y, Xu J, Su T, Liu G, Ren D (2008) Ethylene signaling is required for the acceleration of cell death induced by the activation of AtMEK5 in Arabidopsis. Cell Res 18:422–432PubMedCrossRefGoogle Scholar
  35. Long JC, Caceres JF (2009) The SR protein family of splicing factors: master regulators of gene expression. Biochem J 417:15–27PubMedCrossRefGoogle Scholar
  36. Lopato S, Gattoni R, Fabini G, Stevenin J, Barta A (1999a) A novel family of plant splicing factors with a Zn knuckle motif: examination of RNA binding and splicing activities. Plant Mol Biol 39:761–773PubMedCrossRefGoogle Scholar
  37. Lopato S, Kalyna M, Dorner S, Kobayashi R, Krainer AR, Barta A (1999b) atSRp30, one of two SF2/ASF-like proteins from Arabidopsis thaliana, regulates splicing of specific plant genes. Genes Dev 13:987–1001PubMedCrossRefGoogle Scholar
  38. Lopato S, Forstner C, Kalyna M, Hilscher J, Langhamme U, Indrapichate K, Lorkovi ZJ, Barta A (2002) Network of interactions of a novel plant-specific Arg/Ser-rich protein, atRSZ33, with atSC35-like splicing factors. J Biol Chem 277:39989–39998PubMedCrossRefGoogle Scholar
  39. Manley JL, Krainer AR (2010) A rational nomenclature for serine/arginine-rich protein splicing factors (SR proteins). Genes Dev 24:1073–1074PubMedCrossRefGoogle Scholar
  40. Merdzhanova G, Edmond V, De Seranno S, Van den Broeck A, Corcos L, Brambilla C et al (2008) E2F1 controls alternative splicing pattern of genes involved in apoptosis through upregulation of the splicing factor SC35. Cell Death Differ 15:1815–1823PubMedCrossRefGoogle Scholar
  41. Nilsen TW, Graveley BR (2010) Expansion of the eukaryotic proteome by alternative splicing. Nature 463:457–463PubMedCrossRefGoogle Scholar
  42. Nilsson J, Grahn M, Wright APH (2011) Proteome-wide evidence for enhanced positive Darwinian selection within intrinsically disordered regions in proteins. Genome Biol 12(7):R65Google Scholar
  43. Palusa SG, Ali GS, Reddy ASN (2007) Alternative splicing of pre-mRNAs of Arabidopsis serine/arginine-rich proteins: regulation by hormones and stresses. Plant J 49:1091–1107PubMedCrossRefGoogle Scholar
  44. Rausin G, Tillemans V, Stankovic N, Hanikenne M, Motte P (2010) Dynamic nucleocytoplasmic shuttling of an arabidopsis SR splicing factor: role of the RNA-binding domains. Plant Physiol 153:273–284PubMedCrossRefGoogle Scholar
  45. Shepard PJ, Hertel KJ (2009) The SR protein family. Genome Biol 10:242. doi: 10.1186/gb-2009-10-10-242 PubMedCrossRefGoogle Scholar
  46. Shomron N, Reznik M, Ast G (2004) Splicing factor hSlu7 contains a unique functional domain required to retain the protein within the nucleus. Mol Biol Cell 15:3782–3795PubMedCrossRefGoogle Scholar
  47. Simpson CG, Manthri S, Raczynska KD, Kalyna M, Lewandowska D, Kusenda B et al (2010) Regulation of plant gene expression by alternative splicing. Biochem Soc Trans 38:667–671PubMedCrossRefGoogle Scholar
  48. Swartz JE, Bor Y-C, Misawa Y, Rekosh D, Hammarskjold M-L (2007) The shuttling SR protein 9G8 plays a role in translation of unspliced mRNA containing a constitutive transport element. J Biol Chem 282:19844–19853PubMedCrossRefGoogle Scholar
  49. Takahashi Y, Nasir KHB, Ito A, Kanzaki H, Matsumura H, Saitoh H et al (2007) A high-throughput screen of cell-death-inducing factors in Nicotiana benthamiana identifies a novel MAPKK that mediates INF1-induced cell death signaling and non-host resistance to Pseudomonas cichorii. Plant J 49:1030–1040PubMedCrossRefGoogle Scholar
  50. Tanabe N, Yoshimura K, Kimura A, Yabuta Y, Shigeoka S (2007) Differential expression of alternatively spliced mRNAs of arabidopsis SR protein homologs, atSR30 and atSR45a, in response to environmental stress. Plant Cell Physiol 48:1036–1049PubMedCrossRefGoogle Scholar
  51. Wang BB, Brendel V (2006) Genomewide comparative analysis of alternative splicing in plants. Proc Natl Acad Sci USA 103:7175–7180PubMedCrossRefGoogle Scholar
  52. Warner JR, McIntosh KB (2009) How common are extraribosomal functions of ribosomal proteins? Mol Cell 34:3–11PubMedCrossRefGoogle Scholar
  53. Xu S, Zhang Z, Jing B, Gannon P, Ding J et al (2011) Transportin-SR is required for proper splicing of resistance genes and plant immunity. PLoS Genet 7(6):e1002159. doi: 10.1371/journal.pgen.1002159 PubMedCrossRefGoogle Scholar
  54. Zhang XC, Gassmann W (2003) RPS4-mediated disease resistance requires the combined presence of RPS4 transcripts with full-length and truncated open reading frames. Plant Cell 15:2333–2342PubMedCrossRefGoogle Scholar
  55. Zhang Y, Dorey S, Swiderski M, Jones JDG (2004) Expression of RPS4 in tobacco induces an AvrRps4-independent HR that requires EDS1, SGT1 and HSP90. Plant J 40:213–224PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  1. 1.Department of Plant SciencesUniversity of HyderabadHyderabadIndia
  2. 2.Department of Medical Biochemistry and BiophysicsUmeå UniversityUmeåSweden

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