Penaeus monodon thioredoxin restores the DNA binding activity of oxidized white spot syndrome virus IE1

Jiun Yan Huang, Wang Jing Liu, Han Ching Wang, Der Yen Lee, Jiann Horng Leu, Hao Ching Wang, Mong Hsun Tsai, Shih Ting Kang, I. Tung Chen, Guang Hsiung Kou, Geen Dong Chang, Chu Fang Lo

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15 Citations (Scopus)

Abstract

Aims: In this study we identified viral gene targets of the important redox regulator thioredoxin (Trx), and explored in depth how Trx interacts with the immediate early gene #1 (IE1) of the white spot syndrome virus (WSSV). Results: In a pull-down assay, we found that recombinant Trx bound to IE1 under oxidizing conditions, and a coimmunoprecipitation assay showed that Trx bound to WSSV IE1 when the transfected cells were subjected to oxidative stress. A pull-down assay with Trx mutants showed that no IE1 binding occurred when cysteine 62 was replaced by serine. Electrophoretic mobility shift assay (EMSA) showed that the DNA binding activity of WSSV IE1 was downregulated under oxidative conditions, and that Penaeus monodon Trx (PmTrx) restored the DNA binding activity of the inactivated, oxidized WSSV IE1. Another EMSA experiment showed that IE1's Cys-X-X-Cys motif and cysteine residue 55 were necessary for DNA binding. Measurement of the ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in WSSV-infected shrimp showed that oxidative stress was significantly increased at 48 h postinfection. The biological significance of Trx was also demonstrated in a double-strand RNA Trx knockdown experiment where suppression of shrimp Trx led to significant decreases in mortality and viral copy numbers. Innovation and Conclusion: WSSV's pathogenicity is enhanced by the virus' use of host Trx to rescue the DNA binding activity of WSSV IE1 under oxidizing conditions. Antioxid. Redox Signal.

Original languageEnglish
Pages (from-to)914-926
Number of pages13
JournalAntioxidants and Redox Signaling
Volume17
Issue number6
DOIs
Publication statusPublished - Sep 15 2012
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Physiology
  • Clinical Biochemistry

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    Huang, J. Y., Liu, W. J., Wang, H. C., Lee, D. Y., Leu, J. H., Wang, H. C., Tsai, M. H., Kang, S. T., Chen, I. T., Kou, G. H., Chang, G. D., & Lo, C. F. (2012). Penaeus monodon thioredoxin restores the DNA binding activity of oxidized white spot syndrome virus IE1. Antioxidants and Redox Signaling, 17(6), 914-926. https://doi.org/10.1089/ars.2011.4264