Glyceraldehyde 3-phosphate dehydrogenase negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA

K. Reddisiva Prasanth, Ying Wen Huang, Ming Ru Liou, Robert Yung Liang Wang, Chung Chi Hu, Ching Hsiu Tsai, Menghsiao Meng, Na Sheng Lin, Yau Heiu Hsu

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Abstract

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 ~35 and ~55 kDa were specifically cross-linked with RNA elements, whereupon the ~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.

Original languageEnglish (US)
Pages (from-to)8829-8840
Number of pages12
JournalJournal of Virology
Volume85
Issue number17
DOIs
StatePublished - Sep 2011
Externally publishedYes

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Bamboo mosaic virus
Satellite RNAs
satellite RNA
Mosaic Viruses
Glyceraldehyde-3-Phosphate Dehydrogenases
glyceraldehyde-3-phosphate dehydrogenase
RNA
3' Untranslated Regions
3' untranslated regions
Electrophoretic Mobility Shift Assay
messenger RNA
viruses
Proteins
proteins
gels
Satellite Viruses
Untranslated RNA
Messenger RNA
Nicotiana benthamiana
Host Specificity

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Prasanth, K. R., Huang, Y. W., Liou, M. R., Wang, R. Y. L., Hu, C. C., Tsai, C. H., ... Hsu, Y. H. (2011). Glyceraldehyde 3-phosphate dehydrogenase negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA. Journal of Virology, 85(17), 8829-8840. https://doi.org/10.1128/JVI.00556-11

Glyceraldehyde 3-phosphate dehydrogenase negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA. / Prasanth, K. Reddisiva; Huang, Ying Wen; Liou, Ming Ru; Wang, Robert Yung Liang; Hu, Chung Chi; Tsai, Ching Hsiu; Meng, Menghsiao; Lin, Na Sheng; Hsu, Yau Heiu.

In: Journal of Virology, Vol. 85, No. 17, 09.2011, p. 8829-8840.

Research output: Contribution to journalArticle

Prasanth, KR, Huang, YW, Liou, MR, Wang, RYL, Hu, CC, Tsai, CH, Meng, M, Lin, NS & Hsu, YH 2011, 'Glyceraldehyde 3-phosphate dehydrogenase negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA', Journal of Virology, vol. 85, no. 17, pp. 8829-8840. https://doi.org/10.1128/JVI.00556-11
Prasanth, K. Reddisiva ; Huang, Ying Wen ; Liou, Ming Ru ; Wang, Robert Yung Liang ; Hu, Chung Chi ; Tsai, Ching Hsiu ; Meng, Menghsiao ; Lin, Na Sheng ; Hsu, Yau Heiu. / Glyceraldehyde 3-phosphate dehydrogenase negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA. In: Journal of Virology. 2011 ; Vol. 85, No. 17. pp. 8829-8840.
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