#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Combining ChIP-chip and Expression Profiling to Model the MoCRZ1 Mediated Circuit for Ca/Calcineurin Signaling in the Rice Blast Fungus


Significant progress has been made in defining the central signaling networks in many organisms, but collectively we know little about the downstream targets of these networks and the genes they regulate. To reconstruct the regulatory circuit of calcineurin signal transduction via MoCRZ1, a Magnaporthe oryzae C2H2 transcription factor activated by calcineurin dephosphorylation, we used a combined approach of chromatin immunoprecipitation - chip (ChIP-chip), coupled with microarray expression studies. One hundred forty genes were identified as being both a direct target of MoCRZ1 and having expression concurrently differentially regulated in a calcium/calcineurin/MoCRZ1 dependent manner. Highly represented were genes involved in calcium signaling, small molecule transport, ion homeostasis, cell wall synthesis/maintenance, and fungal virulence. Of particular note, genes involved in vesicle mediated secretion necessary for establishing host associations, were also found. MoCRZ1 itself was a target, suggesting a previously unreported autoregulation control point. The data also implicated a previously unreported feedback regulation mechanism of calcineurin activity. We propose that calcium/calcineurin regulated signal transduction circuits controlling development and pathogenicity manifest through multiple layers of regulation. We present results from the ChIP-chip and expression analysis along with a refined model of calcium/calcineurin signaling in this important plant pathogen.


Vyšlo v časopise: Combining ChIP-chip and Expression Profiling to Model the MoCRZ1 Mediated Circuit for Ca/Calcineurin Signaling in the Rice Blast Fungus. PLoS Pathog 6(5): e32767. doi:10.1371/journal.ppat.1000909
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000909

Souhrn

Significant progress has been made in defining the central signaling networks in many organisms, but collectively we know little about the downstream targets of these networks and the genes they regulate. To reconstruct the regulatory circuit of calcineurin signal transduction via MoCRZ1, a Magnaporthe oryzae C2H2 transcription factor activated by calcineurin dephosphorylation, we used a combined approach of chromatin immunoprecipitation - chip (ChIP-chip), coupled with microarray expression studies. One hundred forty genes were identified as being both a direct target of MoCRZ1 and having expression concurrently differentially regulated in a calcium/calcineurin/MoCRZ1 dependent manner. Highly represented were genes involved in calcium signaling, small molecule transport, ion homeostasis, cell wall synthesis/maintenance, and fungal virulence. Of particular note, genes involved in vesicle mediated secretion necessary for establishing host associations, were also found. MoCRZ1 itself was a target, suggesting a previously unreported autoregulation control point. The data also implicated a previously unreported feedback regulation mechanism of calcineurin activity. We propose that calcium/calcineurin regulated signal transduction circuits controlling development and pathogenicity manifest through multiple layers of regulation. We present results from the ChIP-chip and expression analysis along with a refined model of calcium/calcineurin signaling in this important plant pathogen.


Zdroje

1. EbboleDJ

2007 Magnaporthe as a model for understanding host-pathogen interactions. Annu Rev Phytopathol 45 437 456

2. KankanalaP

CzymmekK

ValentB

2007 Roles for rice membrane dynamics and plasmodesmata during biotrophic invasion by the blast fungus. Plant Cell 19 706 724

3. WilsonRA

TalbotNJ

2009 Under pressure: investigating the biology of plant infection by Magnaporthe oryzae. Nat Rev Microbiol 7 185 195

4. XuJR

ZhaoX

DeanRA

2007 From genes to genomes: a new paradigm for studying fungal pathogenesis in Magnaporthe oryzae. Adv Genet 57 175 218

5. ChoiJ

KimY

KimS

ParkJ

LeeYH

2009 MoCRZ1, a gene encoding a calcineurin-responsive transcription factor, regulates fungal growth and pathogenicity of Magnaporthe oryzae. Fungal Genet Biol 46 243 254

6. ChoiJH

KimY

LeeYH

2009 Functional analysis of MCNA, a gene encoding a catalytic subunit of calcineurin, in the rice blast fungus Magnaporthe oryzae. J Microbiol Biotechnol 19 11 16

7. LeeSC

LeeYH

1998 Calcium/calmodulin-dependent signaling for appressorium formation in the plant pathogenic fungus Magnaporthe grisea. Mol Cells 8 698 704

8. NguyenQB

KadotaniN

KasaharaS

TosaY

MayamaS

2008 Systematic functional analysis of calcium-signalling proteins in the genome of the rice-blast fungus, Magnaporthe oryzae, using a high-throughput RNA-silencing system. Mol Microbiol 68 1348 1365

9. RhoHS

JeonJ

LeeYH

2009 Phospholipase C-mediated calcium signalling is required for fungal development and pathogenicity in Magnaporthe oryzae. Mol Plant Pathol 10 337 346

10. SteinbachWJ

ReedyJL

CramerRAJr

PerfectJR

HeitmanJ

2007 Harnessing calcineurin as a novel anti-infective agent against invasive fungal infections. Nat Rev Microbiol 5 418 430

11. CarafoliE

2002 Calcium signaling: a tale for all seasons. Proc Natl Acad Sci U S A 99 1115 1122

12. MiyakawaT

MizunumaM

2007 Physiological roles of calcineurin in Saccharomyces cerevisiae with special emphasis on its roles in G2/M cell-cycle regulation. Biosci Biotechnol Biochem 71 633 645

13. BaderT

BodendorferB

SchroppelK

MorschhauserJ

2003 Calcineurin is essential for virulence in Candida albicans. Infect Immun 71 5344 5354

14. BlankenshipJR

WormleyFL

BoyceMK

SchellWA

FillerSG

2003 Calcineurin is essential for Candida albicans survival in serum and virulence. Eukaryot Cell 2 422 430

15. OdomA

MuirS

LimE

ToffalettiDL

PerfectJ

1997 Calcineurin is required for virulence of Cryptococcus neoformans. EMBO J 16 2576 2589

16. SanglardD

IscherF

MarchettiO

EntenzaJ

BilleJ

2003 Calcineurin A of Candida albicans: involvement in antifungal tolerance, cell morphogenesis and virulence. Mol Microbiol 48 959 976

17. CamposCB

Di BenedetteJP

MoraisFV

OvalleR

NobregaMP

2008 Evidence for the role of calcineurin in morphogenesis and calcium homeostasis during mycelium-to-yeast dimorphism of Paracoccidioides brasiliensis. Eukaryot Cell 7 1856 1864

18. da Silva FerreiraME

HeinekampT

HartlA

BrakhageAA

SemighiniCP

2007 Functional characterization of the Aspergillus fumigatus calcineurin. Fungal Genet Biol 44 219 230

19. SteinbachWJ

CramerRAJr

PerfectBZ

AsfawYG

SauerTC

2006 Calcineurin controls growth, morphology, and pathogenicity in Aspergillus fumigatus. Eukaryot Cell 5 1091 1103

20. KotheGO

FreeSJ

1998 Calcineurin subunit B is required for normal vegetative growth in Neurospora crassa. Fungal Genet Biol 23 248 258

21. ProkischH

YardenO

DiemingerM

TropschugM

BarthelmessIB

1997 Impairment of calcineurin function in Neurospora crassa reveals its essential role in hyphal growth, morphology and maintenance of the apical Ca2+ gradient. Mol Gen Genet 256 104 114

22. RasmussenC

GarenC

BriningS

KincaidRL

MeansRL

1994 The calmodulin-dependent protein phosphatase catalytic subunit (calcineurin A) is an essential gene in Aspergillus nidulans. EMBO J 13 2545 2552

23. JuvvadiPR

KurokiY

AriokaM

NakajimaH

KitamotoK

2003 Functional analysis of the calcineurin-encoding gene cnaA from Aspergillus oryzae: evidence for its putative role in stress adaptation. Arch Microbiol 179 416 422

24. ViaudMC

BalhaderePV

TalbotNJ

2002 A Magnaporthe grisea cyclophilin acts as a virulence determinant during plant infection. Plant Cell 14 917 930

25. SchumacherJ

de LarrinoaIF

TudzynskiB

2008 Calcineurin-responsive zinc finger transcription factor CRZ1 of Botrytis cinerea is required for growth, development, and full virulence on bean plants. Eukaryot Cell 7 584 601

26. SorianiFM

MalavaziI

da Silva FerreiraME

SavoldiM

Von Zeska KressMR

2008 Functional characterization of the Aspergillus fumigatus CRZ1 homologue, CrzA. Mol Microbiol 67 1274 1291

27. StathopoulosAM

CyertMS

1997 Calcineurin acts through the CRZ1/TCN1-encoded transcription factor to regulate gene expression in yeast. Genes Dev 11 3432 3444

28. KarababaM

ValentinoE

PardiniG

CosteAT

BilleJ

2006 CRZ1, a target of the calcineurin pathway in Candida albicans. Mol Microbiol 59 1429 1451

29. CramerRAJr

PerfectBZ

PinchaiN

ParkS

PerlinDS

2008 Calcineurin target CrzA regulates conidial germination, hyphal growth, and pathogenesis of Aspergillus fumigatus. Eukaryot Cell 7 1085 1097

30. YoshimotoH

SaltsmanK

GaschAP

LiHX

OgawaN

2002 Genome-wide analysis of gene expression regulated by the calcineurin/Crz1p signaling pathway in Saccharomyces cerevisiae. J Biol Chem 277 31079 31088

31. ParkJ

ParkB

JungK

JangS

YuK

2008 CFGP: a web-based, comparative fungal genomics platform. Nucleic Acids Res 36 D562 571

32. CyertMS

2003 Calcineurin signaling in Saccharomyces cerevisiae: how yeast go crazy in response to stress. Biochem Biophys Res Commun 311 1143 1150

33. YiM

ChiMH

KhangCH

ParkSY

KangS

2009 The ER chaperone LHS1 is involved in asexual development and rice infection by the blast fungus Magnaporthe oryzae. Plant Cell 21 681 695

34. GilbertMJ

ThorntonCR

WakleyGE

TalbotNJ

2006 A P-type ATPase required for rice blast disease and induction of host resistance. Nature 440 535 539

35. WinnenburgR

BaldwinTK

UrbanM

RawlingsC

KohlerJ

2006 PHI-base: a new database for pathogen host interactions. Nucleic Acids Res 34 D459 464

36. WinnenburgR

UrbanM

BeachamA

BaldwinTK

HollandS

2008 PHI-base update: additions to the pathogen host interaction database. Nucleic Acids Res 36 D572 576

37. JeonJ

ParkSY

ChiMH

ChoiJ

ParkJ

2007 Genome-wide functional analysis of pathogenicity genes in the rice blast fungus. Nat Genet 39 561 565

38. BalhaderePV

TalbotNJ

2001 PDE1 encodes a P-type ATPase involved in appressorium-mediated plant infection by the rice blast fungus Magnaporthe grisea. Plant Cell 13 1987 2004

39. BaileyTL

ElkanC

1994 Fitting a mixture model by expectation maximization to discover motifs in biopolymers. Proc Int Conf Intell Syst Mol Biol 2 28 36

40. LiuXS

BrutlagDL

LiuJS

2002 An algorithm for finding protein-DNA binding sites with applications to chromatin-immunoprecipitation microarray experiments. Nat Biotechnol 20 835 839

41. GuptaS

StamatoyannopoulosJA

BaileyTL

NobleWS

2007 Quantifying similarity between motifs. Genome Biol 8 R24

42. ZhangH

ZhaoQ

LiuK

ZhangZ

WangY

2009 MgCRZ1, a transcription factor of Magnaporthe grisea, controls growth, development and is involved in full virulence. FEMS Microbiol Lett 293 160 169

43. SorianiFM

MalavaziI

SavoldiM

EspesoE

DinamarcoTM

2010 Identification of possible targets of the Aspergillus fumigatus CRZ1 homologue, CrzA. BMC Microbiol 10 12

44. KrausPR

HeitmanJ

2003 Coping with stress: calmodulin and calcineurin in model and pathogenic fungi. Biochem Biophys Res Commun 311 1151 1157

45. KumarKS

Ravi KumarB

SiddavattamD

SubramanyamC

2006 Characterization of calcineurin-dependent response element binding protein and its involvement in copper-metallothionein gene expression in Neurospora. Biochem Biophys Res Commun 345 1010 1013

46. DengL

SugiuraR

TakeuchiM

SuzukiM

EbinaH

2006 Real-time monitoring of calcineurin activity in living cells: evidence for two distinct Ca2+-dependent pathways in fission yeast. Mol Biol Cell 17 4790 4800

47. SpielvogelA

FindonH

ArstHN

Araujo-BazanL

Hernandez-OrtizP

2008 Two zinc finger transcription factors, CrzA and SltA, are involved in cation homoeostasis and detoxification in Aspergillus nidulans. Biochem J 414 419 429

48. KingsburyTJ

CunninghamKW

2000 A conserved family of calcineurin regulators. Genes Dev 14 1595 1604

49. HiliotiZ

GallagherDA

Low-NamST

RamaswamyP

GajerP

2004 GSK-3 kinases enhance calcineurin signaling by phosphorylation of RCNs. Genes Dev 18 35 47

50. KishiT

IkedaA

NagaoR

KoyamaN

2007 The SCFCdc4 ubiquitin ligase regulates calcineurin signaling through degradation of phosphorylated Rcn1, an inhibitor of calcineurin. Proc Natl Acad Sci U S A 104 17418 17423

51. PinchaiN

PerfectBZ

JuvvadiPR

FortwendelJR

CramerRAJr

2009 Aspergillus fumigatus calcipressin CbpA is involved in hyphal growth and calcium homeostasis. Eukaryot Cell 8 511 519

52. FoxDS

CoxGM

HeitmanJ

2003 Phospholipid-binding protein Cts1 controls septation and functions coordinately with calcineurin in Cryptococcus neoformans. Eukaryot Cell 2 1025 1035

53. ChoquerM

BoccaraM

GoncalvesIR

SoulieMC

Vidal-CrosA

2004 Survey of the Botrytis cinerea chitin synthase multigenic family through the analysis of six euascomycetes genomes. Eur J Biochem 271 2153 2164

54. WernerS

SuguiJA

SteinbergG

DeisingHB

2007 A chitin synthase with a myosin-like motor domain is essential for hyphal growth, appressorium differentiation, and pathogenicity of the maize anthracnose fungus Colletotrichum graminicola. Mol Plant Microbe Interact 20 1555 1567

55. Martin-UrdirozM

RonceroMI

Gonzalez-ReyesJA

Ruiz-RoldanC

2008 ChsVb, a class VII chitin synthase involved in septation, is critical for pathogenicity in Fusarium oxysporum. Eukaryot Cell 7 112 121

56. SoulieMC

PerinoC

PiffeteauA

ChoquerM

MalfattiP

2006 Botrytis cinerea virulence is drastically reduced after disruption of chitin synthase class III gene (Bcchs3a). Cell Microbiol 8 1310 1321

57. BatesS

MacCallumDM

BertramG

MunroCA

HughesHB

2005 Candida albicans Pmr1p, a secretory pathway P-type Ca2+/Mn2+-ATPase, is required for glycosylation and virulence. J Biol Chem 280 23408 23415

58. Ramos-PamplonaM

NaqviNI

2006 Host invasion during rice-blast disease requires carnitine-dependent transport of peroxisomal acetyl-CoA. Mol Microbiol 61 61 75

59. KimS

AhnIP

RhoHS

LeeYH

2005 MHP1, a Magnaporthe grisea hydrophobin gene, is required for fungal development and plant colonization. Mol Microbiol 57 1224 1237

60. MorohashiK

ZhaoM

YangM

ReadB

LloydA

2007 Participation of the Arabidopsis bHLH factor GL3 in trichome initiation regulatory events. Plant Physiol 145 736 746

61. MorohashiK

GrotewoldE

2009 A systems approach reveals regulatory circuitry for Arabidopsis trichome initiation by the GL3 and GL1 selectors. PLoS Genet 5 e1000396

62. CrooksGE

HonG

ChandoniaJM

BrennerSE

2004 WebLogo: a sequence logo generator. Genome Res 14 1188 1190

63. FrangioniJV

NeelBG

1993 Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins. Anal Biochem 210 179 187

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2010 Číslo 5
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#