Open AccessThe 3′‐terminal sequence of Bamboo mosaic virus minus‐strand RNA interacts with RNA‐dependent RNA polymerase and initiates plus‐strand RNA synthesis
Author(s) -
CHEN IHSUAN,
LIN JENWEN,
CHEN YIJING,
WANG ZICHAO,
LIANG LIFANG,
MENG MENGHSIAO,
HSU YAUHEIU,
TSAI CHINGHSIU
Publication year - 2010
Publication title -
molecular plant pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.945
H-Index - 103
eISSN - 1364-3703
pISSN - 1464-6722
DOI - 10.1111/j.1364-3703.2009.00597.x
Subject(s) - biology , rna , rna dependent rna polymerase , rna polymerase , rna editing , polymerase , virology , sequence (biology) , non coding rna , small nuclear rna , microbiology and biotechnology , genetics , gene
SUMMARY 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 .