Alternative system for detection and mapping of activation domains.
The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vectors that allow the fusion of putative activation regions in any reading frame were constructed using Zd. These vectors were used to demonstrate the activity of different classes of activation domains using a chloramphenicol acetyltransferase (cat) reporter gene construct containing seven ZEBRA response elements (Z7). The Zd/Z7 system effectively detected proline-rich, glutamine-rich and acidic activation domains in a variety of cell lines and cell types. Using a bioassay unique to the EBV Zd/Z7 system, fusion constructs can also be tested for the ability to activate gene expression directly from a chromatin structure, the EBV genome. These studies indicate that the Zd/Z7 system is an alternative to GAL4 and can be a useful tool for identifying heterologous activation domains.
Asković
S
S
University of Mississippi Medical Center, Jackson, MS, USA. yar@fiona.umsmed.edu
Baumann
R
R
D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002874:Chromosome Mapping; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D019281:Dimerization; D018274:Electroporation; D017353:Gene Deletion; D015870:Gene Expression; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000737:chemistry; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D014764:Viral Proteins
C063676:BZLF1 protein, Herpesvirus 4, Human; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase