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Arch Virol ; 152(10): 1819-28, 2007.
Article in English | MEDLINE | ID: mdl-17653621

ABSTRACT

We earlier documented the involvement of novel Sp-family-like protein factors in transcription from the Autographa californica nucleopolyhedrovirus (AcNPV) polyhedrin (polh) gene promoter [Ramachandran et al. (2001) J. Biol. Chem. 276: 23440-23449]. These zinc-dependent Sp-like factors bind to two putative Sp-factor-binding motifs, present within the AcSp sequence upstream of the polh promoter, with very high affinity (K(d) = 2.1 x 10(-12) M). Like other polh-promoter-associated host transcription factors, these Sp-like factors display tolerance to high ion concentrations up to even 3 M NaCl. An electrophoretic mobility shift assay demonstrated a probable cross-talk between the Spodoptera frugiperda (Sf9) Sp-family-like proteins and the TFIID complex. In complementary experiments, specific replacements of the Sp-factor-binding motifs with TATA-like elements resulted in expression of a luciferase reporter gene to almost the same level as that obtained with a wild-type native construct. Our results point to the possibility of the involvement of TFIID and Sf9 Sp protein interaction in transcription from the baculovirus polyhedrin promoter.


Subject(s)
Insect Proteins , Nucleopolyhedroviruses/genetics , Sp1 Transcription Factor/physiology , Spodoptera/virology , Viral Proteins/genetics , Animals , Base Sequence , Binding Sites , Cell Extracts/analysis , Cells, Cultured , Cloning, Molecular , Cross-Linking Reagents/metabolism , DNA, Viral/metabolism , Dose-Response Relationship, Drug , Electrophoretic Mobility Shift Assay , Genes, Reporter , Genes, Viral , Genetic Vectors , Luciferases/metabolism , Molecular Sequence Data , Mutation , Nuclear Proteins/metabolism , Occlusion Body Matrix Proteins , Plasmids , Protein Binding , Sodium Chloride/pharmacology , Spodoptera/cytology , Spodoptera/metabolism , Ultraviolet Rays , Viral Structural Proteins/metabolism
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