Identification and zinc dependence of the bovine herpesvirus 1 transactivator protein BICP0.
Bovine herpesvirus 1 (BHV-1) specifies and unspliced early 2.6-kb RNA (ER2.6) which is 3' coterminal with exon 2 of the 2.9-kb immediate-early (IE) RNA. The two transcripts have a common open reading frame (676 codons). The predicted protein, designated BHV-1 infected cell protein 0 (BICP0), contains a zinc finger domain with homology to ICP0 of herpes simplex virus type 1 and protein 61 of varicella-zoster virus, and depending on the promoter, it acts as a strong activator or as a repressor in transient expression assays. In situ immunoadsorbent assays using antisera against synthetic oligopeptides demonstrated that BICP0 accumulates in nuclei of BHV-1-infected cells, as expected for an IE gene product involved in gene regulation. Western blots (immunoblots) revealed a BHV-1-specific 97-kDa protein which was detectable during the IE phase and also at later periods of infection, indicating that the kinetics of BICP0 synthesis is consistent with the switch from IER2.9 to ER2.6. To confirm that ER2.6 encoded the 97-kDa BICP0 protein, a DNA fragment containing BICP0-coding sequences was inserted into the Autographa californica baculovirus genome. A recombinant protein, identified by its reactivity with antipeptide sera, exhibited the same electrophoretic mobility as BICP0 specified by BHV-1. We microinjected Xenopus oocytes with a BICP0 effector plasmid and a promoter-chloramphenicol acetyltransferase plasmid. BICP0-induced stimulation of this promoter was strongly reduced when intracellular zinc was chelated by thionein, indicating that the effect of BICP0 is zinc dependent.
Fraefel
C
C
Institute of Virology, Faculty of Veterinary Medicine, University of Zürich, Switzerland.
Zeng
J
J
Choffat
Y
Y
Engels
M
M
Schwyzer
M
M
Ackermann
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral; D016367:Baculoviridae / Q000235:genetics; D002417:Cattle; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D004880:Ergothioneine / Q000494:pharmacology; D007242:Herpesvirus 1, Bovine / Q000378:metabolism*; D008969:Molecular Sequence Data; D009036:Moths; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000187:drug effects / Q000276:immunology; D015534:Trans-Activators / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D015533:Transcriptional Activation; D044767:Ubiquitin-Protein Ligases; D014764:Viral Proteins / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D014981:Xenopus / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology*
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D004880:Ergothioneine; D044767:Ubiquitin-Protein Ligases; C086729:bICP0 protein, Bovine herpesvirus 1; D015032:Zinc