#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

The RNA Silencing Enzyme RNA Polymerase V Is Required for Plant Immunity


RNA–directed DNA methylation (RdDM) is an epigenetic control mechanism driven by small interfering RNAs (siRNAs) that influence gene function. In plants, little is known of the involvement of the RdDM pathway in regulating traits related to immune responses. In a genetic screen designed to reveal factors regulating immunity in Arabidopsis thaliana, we identified NRPD2 as the OVEREXPRESSOR OF CATIONIC PEROXIDASE 1 (OCP1). NRPD2 encodes the second largest subunit of the plant-specific RNA Polymerases IV and V (Pol IV and Pol V), which are crucial for the RdDM pathway. The ocp1 and nrpd2 mutants showed increases in disease susceptibility when confronted with the necrotrophic fungal pathogens Botrytis cinerea and Plectosphaerella cucumerina. Studies were extended to other mutants affected in different steps of the RdDM pathway, such as nrpd1, nrpe1, ago4, drd1, rdr2, and drm1drm2 mutants. Our results indicate that all the mutants studied, with the exception of nrpd1, phenocopy the nrpd2 mutants; and they suggest that, while Pol V complex is required for plant immunity, Pol IV appears dispensable. Moreover, Pol V defective mutants, but not Pol IV mutants, show enhanced disease resistance towards the bacterial pathogen Pseudomonas syringae DC3000. Interestingly, salicylic acid (SA)–mediated defenses effective against PsDC3000 are enhanced in Pol V defective mutants, whereas jasmonic acid (JA)–mediated defenses that protect against fungi are reduced. Chromatin immunoprecipitation analysis revealed that, through differential histone modifications, SA–related defense genes are poised for enhanced activation in Pol V defective mutants and provide clues for understanding the regulation of gene priming during defense. Our results highlight the importance of epigenetic control as an additional layer of complexity in the regulation of plant immunity and point towards multiple components of the RdDM pathway being involved in plant immunity based on genetic evidence, but whether this is a direct or indirect effect on disease-related genes is unclear.


Vyšlo v časopise: The RNA Silencing Enzyme RNA Polymerase V Is Required for Plant Immunity. PLoS Genet 7(12): e32767. doi:10.1371/journal.pgen.1002434
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002434

Souhrn

RNA–directed DNA methylation (RdDM) is an epigenetic control mechanism driven by small interfering RNAs (siRNAs) that influence gene function. In plants, little is known of the involvement of the RdDM pathway in regulating traits related to immune responses. In a genetic screen designed to reveal factors regulating immunity in Arabidopsis thaliana, we identified NRPD2 as the OVEREXPRESSOR OF CATIONIC PEROXIDASE 1 (OCP1). NRPD2 encodes the second largest subunit of the plant-specific RNA Polymerases IV and V (Pol IV and Pol V), which are crucial for the RdDM pathway. The ocp1 and nrpd2 mutants showed increases in disease susceptibility when confronted with the necrotrophic fungal pathogens Botrytis cinerea and Plectosphaerella cucumerina. Studies were extended to other mutants affected in different steps of the RdDM pathway, such as nrpd1, nrpe1, ago4, drd1, rdr2, and drm1drm2 mutants. Our results indicate that all the mutants studied, with the exception of nrpd1, phenocopy the nrpd2 mutants; and they suggest that, while Pol V complex is required for plant immunity, Pol IV appears dispensable. Moreover, Pol V defective mutants, but not Pol IV mutants, show enhanced disease resistance towards the bacterial pathogen Pseudomonas syringae DC3000. Interestingly, salicylic acid (SA)–mediated defenses effective against PsDC3000 are enhanced in Pol V defective mutants, whereas jasmonic acid (JA)–mediated defenses that protect against fungi are reduced. Chromatin immunoprecipitation analysis revealed that, through differential histone modifications, SA–related defense genes are poised for enhanced activation in Pol V defective mutants and provide clues for understanding the regulation of gene priming during defense. Our results highlight the importance of epigenetic control as an additional layer of complexity in the regulation of plant immunity and point towards multiple components of the RdDM pathway being involved in plant immunity based on genetic evidence, but whether this is a direct or indirect effect on disease-related genes is unclear.


Zdroje

1. MatzkeMKannoTDaxingerLHuettelBMatzkeAJ 2009 RNA-mediated chromatin-based silencing in plants. Curr Opin Cell Biol [doi:10.1016/j.ceb.2009.01.025]

2. LawJAJacobsenSE 2010 Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nature Rev Gent 11 204 220

3. OnoderaYHaagJRReamTNunesPCPontesO 2005 Plant nuclear RNA polymerase IV mediates siRNA and DNA methylation-dependent heterochromatin formation. Cell 120 613 622

4. PontierDYahubyanGVegaDBulskiASaez-VasquezJ 2005 Reinforcement of silencing at transposons and highly repeated sequences requires the concerted action of two distinct RNA polymerases IV in Arabidopsis. Genes Dev 19 2030 2040

5. ReamTSHaagJRWierzbickiATNicoraCDNorbeck 2009 Subunit compositions of the RNA-silencing enzymes Pol IV and Pol V reveal their origins as specialized forms of RNA Polymerase II. Mol Cell 33 192 203

6. ChanSWZilbermanDXieZJohansenLKCarringtonJC 2004 RNA silencing genes control de novo DNA methylation. Science 303 1336

7. XieZJohansenLKGustafsonAMKasschauKDLellisAD 2004 Genetic and functional diversification of small RNA pathways in plants. PLoS Biol 2 e104 doi:10.1371/journal.pbio.0020104

8. ZilbermanDCaoXJacobsenSE 2003 ARGONAUTE4 control of locus-specific siRNA accumulation and DNA and histone methylation. Science 299 716 719

9. ZilbermanDCaoXJohansenLKXieZCarringtonJC 2004 Role of Arabidopsis ARGONAUTE4 in RNA-directed DNA methylation triggered by inverted repeats. Curr Biol 14 1214 1220

10. QiYHeXWangXJKohanyOJurkaJ 2006 Distinct catalytic and non-catalytic roles of ARGONAUTE4 in RNA-directed DNA methylation. Nature 443 1008 1012

11. ChinnusamyVZhuJK 2009 Epigenetic regulation of stress responses in plants. Curr Opin Plant Biol 12 133 139

12. Katiyar-AgarwalSJinH 2010 Role of Small RNAs in Host-Microbe Interactions. Annu Rev Phytopathol 48 225 46

13. NavarroLDunoyerPJayFArnoldBDharmasiriN 2006 A plant miRNA contributes to antibacterial resistance by repressing auxin signaling. Science 312 436 439

14. FahlgrenNHowellMDKasschauKDChapmanEJSullivanCM 2007 High-throughput sequencing of Arabidopsis microRNAs: evidence for frequent birth and death of MIRNA genes. PLoS ONE 2 e219 doi:10.1371/journal.pone.0000219

15. NavarroLJayFNomuraKHeSYVoinnetO 2008 Suppression of the microRNA pathway by bacterial effector proteins. Science 321 964 967

16. LuSSunYHAmersonHChiangVL 2007 MicroRNAs in loblolly pine (Pinus taeda L.) and their association with fusiform rust gall development. Plant J 51 1077 1098

17. HeX-FFangY-YFengLGuoH-S 2008 Characterization of conserved and novel microRNAs and their targets, including aTuMV-induced TIR-NBS-LRR class R gene-derived novel miRNA in Brassica. FEBS Lett 582 2445 2452

18. Ruiz-FerrerVVoinnetO 2009 Roles of plant small RNAs in biotic stress responses. Annu Rev Plant Biol 60 485 510

19. Katiyar-AgarwalSGaoSVivian-SmithAJinH 2007 A novel class of bacteria-induced small RNAs in Arabidopsis. Genes Dev 21 3123 3134

20. Katiyar-AgarwalSMorganRDahlbeckDBorsaniOVillegasAJr 2006 A pathogen-inducible endogenous siRNA in plant immunity. Proc Natl Acad Sci USA 103 18002 18007

21. YiHRichardsEJ 2007 A cluster of disease resistance genes in Arabidopsis is coordinately regulated by transcriptional activation and RNA silencing. Plant Cell 19 2929 2939

22. CoegoARamírezVGilMJFlorsVMauch-ManiB 2005 An Arabidopsis homeodomain transcription factor, OVEREXPRESSOR OF CATIONIC PEROXIDASE 3, mediates resistance to infection by necrotrophic pathogens. Plant Cell 17 2123 2137

23. AgorioAVeraP 2007 ARGONAUTE4 is required for resistance to Pseudomonas syringae in Arabidopsis. Plant Cell 19 3778 3790

24. CoegoARamírezVEllulPMaydaEVeraP 2005 The H2O2-regulated Ep5C gene encodes a peroxidase required for bacterial speck susceptibility in tomato. Plant J 42 283 293

25. RamírezVCoegoALópezAAgorioAFlorsV 2009 Drought tolerance in Arabidopsis is controlled by the OCP3 disease resistance regulator. Plant J 58 578 591

26. RamírezVVan der EntSGarcía-AndradeJCoegoAPieterseCM 2010 OCP3 is an important modulator of NPR1-mediated jasmonic acid-dependent induced defenses in Arabidopsis. Plant Biol 10 199

27. CaoHGlazebrookJClarkeJDVolkoSDongX 1997 The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. Cell 88 57 63

28. HaagJRPontesOPikaardCS 2009 Metal A and metal B sites of nuclear RNA polymerase Pol IV and Pol V are required for siRNA-dependent DNA methylation and gene silencing. PLoS ONE 4 e4110 doi:10.1371/journal.pone.0004110

29. GaoZLiuH-LDaxingerLPontesOHeX 2010 An RNA polymerase II- and AGO4-associated protein acts in RNA-directed DNA methylation. Nature 465 106 109

30. LawJAAusinIJohnsonLMVashishtAAZhuJ-K 2010 A protein complex required for Polymerase V transcripts and RNA directed DNA methylation in Arabidopsis. Curr Biol 20 951 956

31. BenderJ 2004 DNA methylation and epigenetics. Annu Rev Plant Biol 55 41 68

32. HamiltonAVoinnetOChappellLBaulcombeD 2002 Two classes of short interfering RNA in RNA silencing. EMBO J 21 4671 4679

33. RushtonPJSomssichIERinglerPShenQJ 2010 WRKY transcription factors. Trends Plant Sci 15 247 258

34. DongJChenCChenZ 2003 Expression profiles of the Arabidopsis WRKY gene superfamily during plant defense response. Plant Mol Biol 51 21 37

35. CaoXJacobsenSE 2002 Locus-specific control of asymmetric and CpNpG methylation by the DRM and CMT3 methyltransferase genes. Proc Natl Acad Sci USA 99 16491 16498

36. WierzbickiATHaagJRPikaardCS 2008 Noncoding transcription by RNA polymerase Pol IVb/Pol V mediates transcriptional silencing of overlapping and adjacent genes. Cell 135 635 648

37. PontesOCosta-NunesPVithayathilPPikaardCS 2009 RNA polymerase V functions in Arabidopsis interphase heterochromatin organization independently of the 24-nt siRNA-directed DNA methylation pathway. Mol Plant 2 700 710

38. DouetJTutoisSTourmenteS 2009 A Pol V – Mediated Silencing, Independent of RNA – Directed DNA Methylation, Applies to 5S rDNA. PLoS Genet 5 e1000690 doi:10.1371/journal.pgen.1000690

39. JaskiewiczMConrathUPeterhänselC 2011 Chromatin modification acts as a memory for systemic acquired resistance in the plant stress response. EMBO Rep 12 50 55

40. PokholokDKHarbisonCTLevineSColeMHannettNM 2005 Genome-wide map of nucleosome acetylation and methylation in yeast. Cell 122 517 27

41. PontvianneFBlevinsTPikaardCS 2010 Arabidopsis histone lysin methyltransferases. Adv Bot Res 1; 53 1 22

42. KannoTKannoYSiegelRMJangMKLenardoMJ 2004 Selective recognition of acetylated histones by bromodomain proteins visualized in living cells. Mol Cell 13 33 43Kanno et al, 2004;

43. VermeulenMMulderKWDenissovSPijnappelWWvan SchaikFM 2007 Selective anchoring of TFIID to nucleosomes by trimethylation of histone H3 lysine 4. Cell 131 58 69

44. DurrantWEDongX 2004 Systemic acquired resistance. Annu Rev Phytopathol 42 185 209

45. KoornneelAPieterseCMJ 2008 Cross Talk in Defense Signaling. Plant Physiol 146 839 844

46. SpoelSHJohnsonJSDongX 2007 Regulation of tradeoffs between plant defenses against pathogens with different lifestyles. Proc Natl Acad Sci USA 104 18842 18847

47. HaringMOffermannSDankerTHorstIPeterhanselC 2007 Chromatin immunoprecipitation: quantitative analysis and data normalization. Plant Methods 2 11

48. BellCJEckerJR 1994 Assignment of 30 microsatellite loci to the linkage map of Arabidopsis. Genomics 19 137 144

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2011 Číslo 12
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#