In vitro and in planta interaction evidence between Nicotiana tabacum thaumatin-like protein 1 (TLP1) and cucumber mosaic virus proteins.
Using a yeast two-hybrid system, we identified a plant cellular factor that interacts with the proteins of the Cucumber mosaic virus (CMV). Initially 14 candidate genes were isolated from Nicotiana tabacum, using a full-length CMV 1a gene as bait. Among the candidate genes, two were encoding thaumatin-like proteins (TLP), and were designated as Nicotiana tabacum thaumatin-like protein 1 (NtTLP1). Consistent with this observation, recombinant GST-NtTLP1 protein, which was expressed and purified in E. coli, bound tightly to CMV 1a in vitro. In planta interaction was also verified via co-immunoprecipitation. Additionally, NtTLP1 specifically interacted with the CMV movement-related proteins, movement protein and coat protein, in yeast. Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that the expression of NtTLP1 increased as the result of CMV inoculation.
Kim
Min Jung
MJ
School of Life Sciences and Biotechnology, Korea University, 1, 5-ga, Anam-dong, 136-701, Seoul, Sungbuk-gu, Republic of Korea.
Ham
Byung-Kook
BK
Kim
Hwa Ran
HR
Lee
In-Ju
IJ
Kim
Young Jin
YJ
Ryu
Ki Hyun
KH
Park
Young In
YI
Paek
Kyung-Hee
KH
D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D015139:Blotting, Southern; D003001:Cloning, Molecular; D017799:Cucumovirus / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005822:Genetic Vectors; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D047468:Immunoprecipitation; D066298:In Vitro Techniques; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D019697:Onions / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D011485:Protein Binding; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism
C092226:ATLP-1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D010940:Plant Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D011092:Polyethylene Glycols; D012313:RNA; D005982:Glutathione Transferase; D001616:beta-Galactosidase; D005978:Glutathione