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|Title:||Sf-PHB2, A new transcription factor, Drives WSSV Ie1 Gene Expression via a 12-bp DNA Element||Authors:||Ma, G.
|Issue Date:||2012||Citation:||Ma, G., Yu, L., Wang, Q., Liu, W., Cui, Y., Kwang, J. (2012). Sf-PHB2, A new transcription factor, Drives WSSV Ie1 Gene Expression via a 12-bp DNA Element. Virology Journal 9 : -. ScholarBank@NUS Repository. https://doi.org/10.1186/1743-422X-9-206||Abstract:||Background: The WSSV immediate early gene ie1 is highly expressed throughout viral infection cycle and may play a central role in initiating viral replication during infection. Results: Here, a detailed characterization of the ie1 promoter was performed using deletion and mutation analyses to elucidate the role of the individual promoter motifs. Three results were obtained: 1) the ie1 promoter is a classical eukaryotic promoter that contains the initiator element (Inr) and TATA box responsible for the basal promoter activity; 2) mutation or truncation of a predicted Sp1 site decreased the level of promoter activity by about 3-fold, indicating that the Sp1 site is an important cis-element of the promoter; and 3) truncation of a 12-bp sequence that resides at -78/-67 of the ie1 promoter decreased the level of promoter activity by about 14-fold, indicating that the 12-bp motif is a critical upstream element of the ie1 promoter for binding of a strong transcription factor to drive the ie1 gene expression in the cells. Further, the 12-bp DNA binding protein was purified from the nuclear proteins of Sf9 cells using DNA affinity chromatography, and was identified as a homologue of the prohibitin2 protein (named as Sf-PHB2) using mass spectrometry. Furthermore, the DNA binding activity of Sf-PHB2 was verified using a super shift analysis. Conclusion: These results support that the Sf-PHB2 is a novel transcription factor that drives WSSV ie1 gene expression by binding to the 12-bp DNA element. © 2012 Ma et al.; licensee BioMed Central Ltd.||Source Title:||Virology Journal||URI:||http://scholarbank.nus.edu.sg/handle/10635/125516||ISSN:||1743422X||DOI:||10.1186/1743-422X-9-206|
|Appears in Collections:||Staff Publications|
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