期刊
PLANT PATHOLOGY JOURNAL
卷 28, 期 1, 页码 75-80出版社
KOREAN SOC PLANT PATHOLOGY
DOI: 10.5423/PPJ.NT.11.2011.0224
关键词
assembly; cis-elements; EMSA; PVX; SL1(+) RNA-CP interaction
资金
- National Research Foundation [20110012328]
- Ministry of Education, Science, and Technology (MEST)
- Vegetable Breeding Research Center through Agriculture Research Center from Ministry for Food, Agriculture, Forestry and Fisheries [710001-03]
- Rural Development Administration [PJ00819801]
Potato virus X (PVX) contains cis-acting elements including stem-loop 1 (SL1) RNA at the 5' region; SL1 is conserved among all potexviruses. The SL1 at the positive-sense RNA, SL1(+), is required for PVX RNA replication, cell-to-cell movement, and translation. Previous research demonstrated that SL1(+) RNA also serves as the origin of assembly for encapsidation of PVX RNA. To identify the essential sequences and/or regions for capsid protein (CP) subunit recognition within SL1(+) RNA, we used electrophoretic mobility shift assays (EMSA), UV cross-linking, and yeast threehybrid analyses. The EMSA and UV cross-linking analyses with PVX CP subunits and RNA transcripts corresponding to the SL1(+) RNA showed that the SL1(+) RNA formed complexes with CP subunits. We also conducted EMSA and yeast three-hybrid analyses with RNAs containing various mutations of SL1(+) RNA elements. These analyses indicated that SL1(+) RNA is required for the interaction with PVX CP and that the RNA sequences located at the loop C and tetra loop of the SL1(+) are crucial for CP binding. These results indicate that, in addition to being important for RNA accumulation, the SL1(+) RNA from the 5' region of the PVX genome is also required for specific binding of PVX CP.
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