2002 Fiscal Year Final Research Report Summary
Elucidation of the mechanisms of tobacco mosaic virus RNA replication
| Project/Area Number |
13460018
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物保護
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
MASAYUKI Ishikawa Graduate School of Agriculture Hokkaido Univ., Associate Professor, 大学院・農学研究科, 助教授 (70192482)
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| Co-Investigator(Kenkyū-buntansha) |
SATOSHI Naito Graduate School of Agriculture Hokkaido Univ., Professor, 大学院・農学研究科, 教授 (20164105)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Tobacco mosaic virus / RNA replication / host factor / Arabidopsis thaliana |
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| Research Abstract |
Arabidopsis TOM1 and TOM2A are necessary for efficient multiplication of Tobacco mosaic virus (TMV). In this study, we have explored the following two points in order to reveal the function of the TOM1 and TOM2A proteins in TMV multiplication. (1) Subcellular localization of TOM1 and TOM2A : Green fluorescent protein (GFP)-tagged TOM1 and TOM2A proteins are found to be targeted to the vacuolar membrane (tonoplast)-like structure in plant cells. In subcellular fractionation analyses, GFP-TOM2A and TOM2A were predominantly fractionated to low-density, tonoplast-rich fractions, whereas TOM1-GFP and TOM1 were distributed mainly into the tonoplast-rich fractions and partially into higher-buoyant-density fractions containing membranes from several other organelles. The TMV-coded replication proteins were cofractionated with both TOM1 and viral RNA-dependent RNA polymerase activity. The replication proteins were also found in the fractions containing non-membrane-bound proteins, but neither TOM1 nor the polymerase activity was detected there. These observations suggest that the formation of TMV RNA replication complex occurs on TOM1-containing membranes. (2) interaction between TOM1 and TOM2A : To examine possible interaction between TOM1 and TOM2A, the split-ubiquitin assay was utilized. This method demonstrated an interaction of TOM2A both with itself and with TOM1. These results suggest that TOM2A is also an integral part of TMV replication complex.
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