A Viral Satellite RNA Induces Yellow Symptoms on Tobacco by Targeting
a Gene Involved in Chlorophyll Biosynthesis using the RNA Silencing
Machinery
Symptoms on virus-infected plants are often very specific to the given virus. The
molecular mechanisms involved in viral symptom induction have been extensively
studied, but are still poorly understood. Cucumber mosaic virus
(CMV) Y satellite RNA (Y-sat) is a non-coding subviral RNA and modifies the
typical symptom induced by CMV in specific hosts; Y-sat causes a bright yellow
mosaic on its natural host Nicotiana tabacum. The Y-sat-induced
yellow mosaic failed to develop in the infected Arabidopsis and
tomato plants suggesting a very specific interaction between Y-sat and its host.
In this study, we revealed that Y-sat produces specific short interfering RNAs
(siRNAs), which interfere with a host gene, thus inducing the specific symptom.
We found that the mRNA of tobacco magnesium protoporphyrin chelatase subunit I
(ChlI, the key gene involved in chlorophyll synthesis) had
a 22-nt sequence that was complementary to the Y-sat sequence, including four
G-U pairs, and that the Y-sat-derived siRNAs in the virus-infected plant
downregulate the mRNA of ChlI by targeting the complementary
sequence. ChlI mRNA was also downregulated in the transgenic
lines that express Y-sat inverted repeats. Strikingly, modifying the Y-sat
sequence in order to restore the 22-nt complementarity to
Arabidopsis and tomato ChlI mRNA resulted
in yellowing symptoms in Y-sat-infected Arabidopsis and tomato,
respectively. In 5′-RACE experiments, the ChlI transcript
was cleaved at the expected middle position of the 22-nt complementary sequence.
In GFP sensor experiments using agroinfiltration, we further demonstrated that
Y-sat specifically targeted the sensor mRNA containing the 22-nt complementary
sequence of ChlI. Our findings provide direct evidence that the
identified siRNAs derived from viral satellite RNA directly modulate the viral
disease symptom by RNA silencing-based regulation of a host gene.
Vyšlo v časopise:
A Viral Satellite RNA Induces Yellow Symptoms on Tobacco by Targeting
a Gene Involved in Chlorophyll Biosynthesis using the RNA Silencing
Machinery. PLoS Pathog 7(5): e32767. doi:10.1371/journal.ppat.1002021
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002021
Souhrn
Symptoms on virus-infected plants are often very specific to the given virus. The
molecular mechanisms involved in viral symptom induction have been extensively
studied, but are still poorly understood. Cucumber mosaic virus
(CMV) Y satellite RNA (Y-sat) is a non-coding subviral RNA and modifies the
typical symptom induced by CMV in specific hosts; Y-sat causes a bright yellow
mosaic on its natural host Nicotiana tabacum. The Y-sat-induced
yellow mosaic failed to develop in the infected Arabidopsis and
tomato plants suggesting a very specific interaction between Y-sat and its host.
In this study, we revealed that Y-sat produces specific short interfering RNAs
(siRNAs), which interfere with a host gene, thus inducing the specific symptom.
We found that the mRNA of tobacco magnesium protoporphyrin chelatase subunit I
(ChlI, the key gene involved in chlorophyll synthesis) had
a 22-nt sequence that was complementary to the Y-sat sequence, including four
G-U pairs, and that the Y-sat-derived siRNAs in the virus-infected plant
downregulate the mRNA of ChlI by targeting the complementary
sequence. ChlI mRNA was also downregulated in the transgenic
lines that express Y-sat inverted repeats. Strikingly, modifying the Y-sat
sequence in order to restore the 22-nt complementarity to
Arabidopsis and tomato ChlI mRNA resulted
in yellowing symptoms in Y-sat-infected Arabidopsis and tomato,
respectively. In 5′-RACE experiments, the ChlI transcript
was cleaved at the expected middle position of the 22-nt complementary sequence.
In GFP sensor experiments using agroinfiltration, we further demonstrated that
Y-sat specifically targeted the sensor mRNA containing the 22-nt complementary
sequence of ChlI. Our findings provide direct evidence that the
identified siRNAs derived from viral satellite RNA directly modulate the viral
disease symptom by RNA silencing-based regulation of a host gene.
Zdroje
1. SimonAERoossinckMJHaveldaZ
2004
Plant virus satellite and defective interfering RNAs: new
paradigms for a new century.
Annu Rev Phytopathol
42
415
437
2. HuCCHsuYHLinNS
2009
Satellite RNAs and satellite viruses of plants.
Viruses
1
1325
1350
3. HuangYWHuCCLinNSHsuYH
2010
Mimicry of molecular pretenders: the terminal structures of
satellites associated with plant RNA viruses.
RNA Biol
7
162
171
4. TakanamiY
1981
A striking change in symptoms on Cucumber mosaic
virus-infected tobacco plants induced by a satellite
RNA.
Virology
109
120
126
5. XuPRoossinckMJ
2000
Cucumber mosaic virus D satellite RNA-induced
programmed cell death in tomato.
Plant Cell
12
1079
1092
6. MasutaCTakanamiY
1989
Determination of sequence and structural requirements for
pathogenicity of a Cucumber mosaic virus satellite RNA
(Y-satRNA).
Plant Cell
1
1165
1173
7. KuwataSMasutaCTakanamiY
1991
Reciprocal phenotype alterations between two satellite RNAs of
Cucumber mosaic virus.
J Gen Virol
72
2385
2389
8. DevicMJaegleMBaulcombeD
1989
Symptom production on tobacco and tomato is determined by two
distinct domains of the satellite RNA of Cucumber mosaic
virus (strain Y).
J Gen Virol
70
2765
2774
9. JaegleMDevicMLongstaffMBaulcombeD
1990
Cucumber mosaic virus satellite RNA (Y strain):
analysis of sequences which affect yellow mosaic symptoms on
tobacco.
J Gen Virol
71
1905
1912
10. SleatDEPalukaitisP
1992
A single nucleotide change within a plant virus satellite RNA
alters the host specificity of disease induction.
Plant J
2
43
49
11. MasutaCSuzukiMKuwataSTakanamiYKoiwaiA
1993
Yellow mosaic symptoms induced by Y satellite RNA of
Cucumber mosaic virus is regulated by a single
incompletely dominant gene in wild Nicotiana
species.
Phytopathology
83
411
413
12. WangMBBianXYWuLMLiuLXSmithNA
2004
On the role of RNA silencing in the pathogenicity and evolution
of viroids and viral satellites.
Proc Natl Acad Sci U S A
101
3275
3280
13. Ruiz-FerrerVVoinnetO
2009
Roles of plant small RNAs in biotic stress
responses.
Annu Rev Plant Biol
60
485
510
14. PhillipsJRDalmayTBartelsD
2007
The role of small RNAs in abiotic stress.
FEBS Lett
581
3592
3597
15. VoinnetO
2009
Origin, biogenesis, and activity of plant
microRNAs.
Cell
136
669
687
16. VaucheretHMalloryACBartelDP
2006
AGO1 homeostasis entails coexpression of MIR168 and AGO1 and
preferential stabilization of miR168 by AGO1.
Mol Cell
22
129
136
17. BrosnanCAVoinnetO
2009
The long and the short of noncoding RNAs.
Curr Opin Cell Biol
21
416
425
18. LlaveC
2004
MicroRNAs: more than a role in plant development?
Mol Plant Pathol
5
361
366
19. DingSWVoinnetO
2007
Antiviral immunity directed by small RNAs.
Cell
130
413
426
20. CsorbaTPantaleoVBurgyánJ
2009
RNA silencing: an antiviral mechanism.
Adv Virus Res
75
35
71
21. MlotshwaSPrussGJVanceV
2008
Small RNAs in viral infection and host defense.
Trends Plant Sci
13
375
382
22. HaveldaZHornyikCValocziABurgyanJ
2005
Defective interfering RNA hinders the activity of a
tombusvirus-encoded posttranscriptional gene silencing
suppressor.
J Virol
79
450
457
23. SzittyaGMolnarASilhavyDHornyikCBurgyánJ
2002
Short defective interfering RNAs of tombusviruses are not
targeted but trigger post-transcriptional gene silencing against their
helper virus.
Plant Cell
14
359
372
24. OmarovRTRezendeJAScholthofHB
2004
Host-specific generation and maintenance of Tomato bushy
stunt virus defective interfering RNAs.
Mol Plant-Microbe Interact
17
195
201
25. MoissiardGVoinnetO
2006
RNA silencing of host transcripts by cauliflower mosaic virus
requires coordinated action of the four Arabidopsis
Dicer-like proteins.
Proc Natl Acad Sci U S A
103
19593
19598
26. QiXBaoFSXieZ
2009
Small RNA deep sequencing reveals role for Arabidopsis
thaliana RNA-dependent RNA polymerases in viral siRNA
biogenesis.
PLoS One
4
e4971
27. MoulinMMcCormacACTerryMJSmithAG
2008
Tetrapyrrole profiling in Arabidopsis seedlings
reveals that retrograde plastid nuclear signaling is not due to
Mg-protoporphyrin IX accumulation.
Proc Natl Acad Sci U S A
105
15178
15183
28. FitzmauriceWPNguyenLVWernsmanEAThompsonWFConklingMA
1999
Transposon tagging of the sulfur gene of tobacco using engineered
maize Ac/Ds elements.
Genetics
153
1919
1928
29. KjemtrupSSampsonKSPeeleCGNguyenLVConklingMA
1998
Gene silencing from plant DNA carried by a
Geminivirus.
Plant J
14
91
100
30. PetersenBOAlbrechtsenM
2005
Evidence implying only unprimed RdRP activity during transitive
gene silencing in plants.
Plant Mol Biol
58
575
583
31. TuttleJRIdrisAMBrownJKHaiglerCHRobertsonD
2008
Geminivirus-mediated gene silencing from
Cotton leaf crumple virus is enhanced by low
temperature in cotton.
Plant Physiol
148
41
50
32. HuangYSLiHM
2009
Arabidopsis CHLI2 can substitute for
CHLI1.
Plant Physiol
150
636
645
33. HaveldaZVarallyayEValocziABurgyánJ
2008
Plant virus infection-induced persistent host gene downregulation
in systemically infected leaves.
Plant J
55
278
288
34. MasutaCKuwataSMatzuzakiTTakanamiYKoiwaiA
1992
A plant virus satellite RNA exhibits a significant sequence
complementarity to a chloroplast tRNA.
Nucleic Acids Res
20
2885
35. MasutaCHayashiYWangWQTakanamiY
1990
Comparison of four satellite RNA isolates of Cucumber
mosaic virus.
Ann Phytopath Soc Japan
56
207
212
36. KosakaYFukunishiT
1997
Multiple inoculation with three attenuated viruses for the
control of cucumber virus disease.
Plant Dis
81
733
738
37. OtagakiSAraiMTakahashiAGotoKHongJS
2006
Rapid induction of transcriptional and post-transcriptional gene
silencing using a novel Cucumber mosaic virus
vector.
Plant Biotechnol
23
259
265
38. MatsuoKHongJSTabayashiNItoAMasutaC
2007
Development of Cucumber mosaic virus as a vector
modifiable for different host species to produce therapeutic
proteins.
Planta
225
277
286
39. KasschauKDXieZAllenELlaveCChapmanEJ
2003
P1/HC-Pro, a viral suppressor of RNA silencing, interferes with
Arabidopsis development and miRNA
function.
Dev Cell
4
205
217
40. ChapmanEJProkhnevskyAIGopinathKDoljaVVCarringtonJC
2004
Viral RNA silencing suppressors inhibit the microRNA pathway at
an intermediate step.
Genes Dev
18
1179
1186
41. VaistijFEJonesLBaulcombeDC
2002
Spreading of RNA targeting and DNA methylation in RNA silencing
requires transcription of the target gene and a putative RNA-dependent RNA
polymerase.
Plant Cell
14
857
867
42. HimberCDunoyerPMoissiardGRitzenthalerCVoinnetO
2003
Transitivity-dependent and -independent cell-to-cell movement of
RNA silencing.
EMBO J
22
4523
4533
43. AllenEXieZGustafsonAMCarringtonJC
2005
MicroRNA-directed phasing during trans-acting siRNA biogenesis in
plants.
Cell
121
207
221
44. DuQSDuanCGZhangZHFangYYFangRX
2007
DCL4 targets Cucumber mosaic virus satellite RNA
at novel secondary structures.
J Virol
81
9142
9151
45. RuizMTVoinnetOBaulcombeDC
1998
Initiation and maintenance of virus-induced gene
silencing.
Plant Cell
10
937
946
46. SchattatMHKlosgenRBMarquesJP
2004
A novel vector for efficient gene silencing in
plants.
Plant Mol Biol Rep
22
145
153
47. SendaMMasutaCOhnishiSGotoKKasaiA
2004
Patterning of virus-infected Glycine max seed
coat is associated with suppression of endogenous silencing of chalcone
synthase genes.
Plant Cell
16
807
818
48. MasutaCTanakaHUeharaKKuwataSKoiwaiA
1995
Broad resistance to plant viruses in transgenic plants conferred
by antisense inhibition of a host gene essential in
S-adenosylmethionine-dependent transmethylation reactions.
Proc Natl Acad Sci U S A
92
6117
6121
49. GotoKKoboriTKosakaYNatsuakiTMasutaC
2007
Characterization of silencing suppressor 2b of Cucumber
mosaic virus based on examination of its small RNA-binding
abilities.
Plant Cell Physiol
48
1050
1060
50. ValocziAHornyikCVargaNBurgyánJKauppinenS
2004
Sensitive and specific detection of microRNAs by northern blot
analysis using LNA-modified oligonucleotide probes.
Nucleic Acids Res
32
e175
51. LlaveCXieZKasschauKDCarringtonJC
2002
Cleavage of Scarecrow-like mRNA targets directed by a class of
Arabidopsis miRNA.
Science
297
2053
2056
52. ShimuraHFukagawaTMeguroAYamadaHOh-HiraM
2008
A strategy for screening an inhibitor of viral silencing
suppressors, which attenuates symptom development of plant
viruses.
FEBS Lett
582
4047
4052
53. LiRLiYKristiansenKWangJ
2008
SOAP: short oligonucleotide alignment program.
Bioinformatics
24
713
714
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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