The 5' Cap of Tobacco Mosaic Virus (TMV) Is Required for Virion Attachment to the Actin/Endoplasmic Reticulum Network During Early Infection

被引:52
作者
Christensen, Nynne [1 ]
Tilsner, Jens [1 ]
Bell, Karen [1 ]
Hammann, Philippe [2 ]
Parton, Richard [3 ]
Lacomme, Christophe [1 ]
Oparka, Karl [1 ]
机构
[1] Univ Edinburgh, Inst Mol Plant Sci, Edinburgh EH9 3JR, Midlothian, Scotland
[2] Inst Biol Mol Plantes, F-67084 Strasbourg, France
[3] Univ Oxford, Dept Biochem, Oxford OX1 3QU, England
基金
英国生物技术与生命科学研究理事会;
关键词
direct fluorescence labelling; live cell imaging; methylguanosine cap; RNA anchoring; tobacco mosaic virus; viral RNA; CELL-TO-CELL; MESSENGER-RNA LOCALIZATION; MOVEMENT PROTEIN; ENDOPLASMIC-RETICULUM; FLUORESCENT PROTEIN; TRANSLATIONAL CONTROL; REPLICATION COMPLEX; GENE-EXPRESSION; EARLY-STAGE; PLANT;
D O I
10.1111/j.1600-0854.2009.00889.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Almost nothing is known of the earliest stages of plant virus infections. To address this, we microinjected Cy3 (UTP)-labelled tobacco mosaic virus (TMV) into living tobacco trichome cells. The Cy3-virions were infectious, and the viral genome trafficked from cell to cell. However, neither the fluorescent vRNA pool nor the co-injected green fluorescent protein (GFP) left the injected trichome, indicating that the synthesis of (unlabelled) progeny viral (v)RNA is required to initiate cell-to-cell movement, and that virus movement is not accompanied by passive plasmodesmatal gating. Cy3-vRNA formed granules that became anchored to the motile cortical actin/endoplasmic reticulum (ER) network within minutes of injection. Granule movement on actin/ER was arrested by actin inhibitors indicating actin-dependent RNA movement. The 5' methylguanosine cap was shown to be required for vRNA anchoring to the actin/ER. TMV vRNA lacking the 5' cap failed to form granules and was degraded in the cytoplasm. Removal of the 3' untranslated region or replicase both inhibited replication but did not prevent granule formation and movement. Dual-labelled TMV virions in which the vRNA and the coat protein were highlighted with different fluorophores showed that both fluorescent signals were initially located on the same ER-bound granules, indicating that TMV virions may become attached to the ER prior to uncoating of the viral genome.
引用
收藏
页码:536 / 551
页数:16
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