Functional analysis of evolutionary conserved clustering of bZIP binding sites in the baculovirus homologous regions (hrs) suggests a cooperativity between host and viral transcription factors.
The genome of the Autographa californica Multinucleocapsid Polyhedrosis Virus (AcMNPV) contains nine interspersed homologous regions (hrs) that function as potent enhancer sequences when linked in cis to either viral or heterologous RNA polymerase II-dependent promoters. Their activity is strongly increased by the binding of the major immediate early viral transregulator IE1 on 28-mer palindromic sites present in hrs. We show that hrs of AcMNPV additionally carry, in the interpalindromic sequences, a large number of cAMP response elements (CRE) and TPA response elements (TRE), known to bind ubiquitous cellular transcription factors of the bZIP family. Moreover, these clusters of CRE and TRE motifs are concentrated in hrs. Analysis of the 25 baculovirus genomes sequenced so far reveals that these motifs are evolutionary conserved in Lepidoptera NPVs, suggesting a functional role in the hr enhancer function. Consistently, EMSA experiments indicate that CRE and on a lesser extent TRE sites specifically bind insect host factors. Moreover, reporter assays reveal that these CRE sites have an additive stimulatory effect on RNAPol II-dependent transcription in Sf9 cells and are potentially able to synergize with the IE1-binding palindrome.
Landais
Igor
I
Laboratoire de Pathologie Comparée, UMR 5087, 30380 Saint Christol-les-Alès, France.
Vincent
Rachel
R
Bouton
Martine
M
Devauchelle
Gérard
G
Duonor-Cerutti
Martine
M
Ogliastro
Mylène
M
D000494:Allosteric Regulation; D000818:Animals; D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002460:Cell Line; D017124:Conserved Sequence; D019143:Evolution, Molecular; D015967:Gene Expression Regulation, Viral; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011485:Protein Binding; D020218:Response Elements / Q000235:genetics
D050976:Basic-Leucine Zipper Transcription Factors