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標題: The 3 '-terminal sequence of Bamboo mosaic virus minus-strand RNA interacts with RNA-dependent RNA polymerase and initiates plus-strand RNA synthesis
作者: Chen, I.H.
Lin, J.W.
Chen, Y.J.
Wang, Z.C.
Liang, L.F.
Meng, M.H.
Hsu, Y.H.
Tsai, C.H.
關鍵字: helicase-like domain;potexvirus rna;functional-analysis;in-vitro;3'-untranslated region;replication;guanylyltransferase;accumulation;proteins;promoter
Project: Molecular Plant Pathology
期刊/報告no:: Molecular Plant Pathology, Volume 11, Issue 2, Page(s) 203-212.
A 3'-terminal, 77-nucleotide sequence of Bamboo mosaic virus (BaMV) minus-strand RNA (Ba-77), comprising a 5' stem-loop, a spacer and a 3'-CUUUU sequence, can be used to initiate plus-strand RNA synthesis in vitro. To understand the mechanism of plus-strand RNA synthesis, mutations were introduced in the 5' untranslated region of BaMV RNA, resulting in changes at the 3' end of minus-strand RNA. The results showed that at least three uridylate residues in 3'-CUUUU are required and the changes at the penultimate U are deleterious to viral accumulation in Nicotiana benthamiana protoplasts. Results from UV-crosslinking and in vitro RNA-dependent RNA polymerase competition assays suggested that the replicase preferentially interacts with the stem structure of Ba-77. Finally, CMV/83 + UUUUC, a heterologus RNA, which possesses about 80 nucleotides containing the 3'-CUUUU pentamer terminus, and which folds into a secondary structure similar to that of Ba-77, could be used as template for RNA production by the BaMV replicase complex in vitro.
ISSN: 1464-6722
DOI: 10.1111/j.1364-3703.2009.00597.x
Appears in Collections:生物科技學研究所

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