Characterization of proteins binding to the ZII element in the Epstein-Barr virus BZLF1 promoter: transactivation by ATF1.
Previous studies have shown that the ZII element in the BZLF1 promoter (P1) is responsive to TPA and anti-immunoglobulin induction. In this report, we have studied the DNA/protein complexes formed when ZII is used as a binding site. Twelve distinct DNA/protein complexes were seen in mobility shift experiments using Akata cell nuclear extracts and radiolabeled ZII. Eleven of these complexes were also formed when either BJAB or Raji cell nuclear extracts were used in the binding reaction. Six DNA/protein complexes were affected by mutations in the core TGACATCA motif of ZII which abolish responsiveness to TPA, anti-immunoglobulin treatment, and HHV6 transactivation. The relative sizes of the proteins in the DNA/protein complexes were determined by UV crosslinking. Four distinct specific binding proteins affected by core mutations in ZII were identified as ATFa, ATF1, ATF2, and c-jun. Overexpression of ATF1 in cotransfection experiments caused transactivation of the wild-type P1 promoter but had no effect on a promoter containing a mutant ZII element. An ATF1 mutant with a deleted DNA binding domain failed to transactivate P1. Overexpression of c-jun, ATFa, or ATF2 had no effect on the wild-type or mutant P1 promoter. Our results suggest that ATF1 interacts with the ZII element and may be involved in Epstein-Barr virus reactivation.
Wang
Y C
YC
The Institute of Biotechnology, Center for Molecular Medicine, University of Texas Health Science Center at San Antonio, 78245, USA.
Huang
J M
JM
Montalvo
E A
EA
D051697:Activating Transcription Factor 1; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000235:genetics*; D017930:Genes, Reporter; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
D051697:Activating Transcription Factor 1; C063676:BZLF1 protein, Herpesvirus 4, Human; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; D013755:Tetradecanoylphorbol Acetate