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標題: Glyceraldehyde 3-Phosphate Dehydrogenase Negatively Regulates the Replication of Bamboo Mosaic Virus and Its Associated Satellite RNA
作者: Prasanth, K.R.
Huang, Y.W.
Liou, M.R.
Wang, R.Y.L.
Hu, C.C.
Tsai, C.H.
Meng, M.H.
Lin, N.S.
Hsu, Y.H.
關鍵字: minus-strand rna;turnip-crinkle-virus;hepatitis-a virus;gene-block;protein-1;stem-loop structure;synthesis in-vitro;genomic rna;nicotiana-benthamiana;nucleotide-sequence;binding-protein
Project: Journal of Virology
期刊/報告no:: Journal of Virology, Volume 85, Issue 17, Page(s) 8829-8840.
The identification of cellular proteins associated with virus replicase complexes is crucial to our understanding of virus-host interactions, influencing the host range, replication, and virulence of viruses. A previous in vitro study has demonstrated that partially purified Bamboo mosaic virus (BaMV) replicase complexes can be employed for the replication of both BaMV genomic and satellite BaMV (satBaMV) RNAs. In this study, we investigated the BaMV and satBaMV 3' untranslated region (UTR) binding proteins associated with these replicase complexes. Two cellular proteins with molecular masses of similar to 35 and similar to 55 kDa were specifically cross-linked with RNA elements, whereupon the similar to 35-kDa protein was identified as the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Gel mobility shift assays confirmed the direct interaction of GAPDH with the 3' UTR sequences, and competition gel shift analysis revealed that GAPDH binds preferentially to the positive-strand BaMV and satBaMV RNAs over the negative-strand RNAs. It was observed that the GAPDH protein binds to the pseudoknot poly(A) tail of BaMV and stem-loop-C poly(A) tail of satBaMV 3' UTR RNAs. It is important to note that knockdown of GAPDH in Nicotiana benthamiana enhances the accumulation of BaMV and satBaMV RNA; conversely, transient overexpression of GAPDH reduces the accumulation of BaMV and satBaMV RNA. The recombinant GAPDH principally inhibits the synthesis of negative-strand RNA in exogenous RdRp assays. These observations support the contention that cytosolic GAPDH participates in the negative regulation of BaMV and satBaMV RNA replication.
ISSN: 0022-538X
DOI: 10.1128/jvi.00556-11
Appears in Collections:生物科技學研究所

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