1. The promoter of the white spot syndrome virus immediate-early gene WSSV108 is activated by the cellular KLF transcription factor.
- Author
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Liu WJ, Lo CF, Kou GH, Leu JH, Lai YJ, Chang LK, and Chang YS
- Subjects
- Amino Acid Sequence, Animals, Arthropod Proteins genetics, Base Sequence, Binding Sites genetics, Blotting, Western, Electrophoretic Mobility Shift Assay, Gene Expression Regulation, Viral, Host-Pathogen Interactions genetics, Immediate-Early Proteins, Kruppel-Like Transcription Factors genetics, Molecular Sequence Data, Penaeidae genetics, Penaeidae metabolism, Penaeidae virology, Protein Binding, RNA Interference, Reverse Transcriptase Polymerase Chain Reaction, Transcriptional Activation, Viral Proteins metabolism, Virus Replication genetics, White spot syndrome virus 1 metabolism, White spot syndrome virus 1 physiology, Arthropod Proteins metabolism, Genes, Immediate-Early genetics, Kruppel-Like Transcription Factors metabolism, Promoter Regions, Genetic genetics, Viral Proteins genetics, White spot syndrome virus 1 genetics
- Abstract
A series of deletion and mutation assays of the white spot syndrome virus (WSSV) immediate-early gene WSSV108 promoter showed that a Krüppel-like factor (KLF) binding site located from -504 to -495 (relative to the transcription start site) is important for the overall level of WSSV108 promoter activity. Electrophoretic mobility shift assays further showed that overexpressed recombinant Penaeus monodon KLF (rPmKLF) formed a specific protein-DNA complex with the (32)P-labeled KLF binding site of the WSSV108 promoter, and that higher levels of Litopenaeus vannamei KLF (LvKLF) were expressed in WSSV-infected shrimp. A transactivation assay indicated that the WSSV108 promoter was strongly activated by rPmKLF in a dose-dependent manner. Lastly, we found that specific silencing of LvKLF expression in vivo by dsRNA injection dramatically reduced both WSSV108 expression and WSSV replication. We conclude that shrimp KLF is important for WSSV genome replication and gene expression, and that it binds to the WSSV108 promoter to enhance the expression of this immediate-early gene., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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