Cellular and viral factors regulate the varicella-zoster virus gE promoter during viral replication.
Varicella-zoster virus (VZV) glycoprotein E (gE) is essential for viral replication and is involved in cell-to-cell spread, secondary envelopment, and entry. We created a set of mutations in the gE promoter to investigate the role of viral and cellular transcriptional factors in regulation of the gE promoter. Deletion or point mutation of the two Sp1 sites in the gE promoter abolished Sp1 binding and IE62-mediated transactivation of the gE promoter in vitro. Incorporation of the deletion or the point mutations disrupting both of the Sp1 binding sites into the VZV genome was not compatible with viral replication. A point mutation altering the atypical Sp1 binding site was lethal, while altering the second site impaired VZV replication significantly, indicating functional differences between the two Sp1 binding sites. Deletions in the gE promoter that abolished putative binding sites for cellular transcriptional factors other than Sp1, identified by bioinformatics analysis, were inserted in the VZV genome. Replication of the viruses with mutations of the gE promoter did not differ from control recombinants in melanoma cells or primary human tonsil T cells in vitro. These deletions did not affect infection of human skin xenografts in SCIDhu mice. These results indicate that Sp1 is required for IE62-mediated transactivation of the gE promoter and that this transcriptional factor appears to be the only cellular factor essential for regulation of the gE promoter.
Berarducci
Barbara
B
Department of Pediatrics, Stanford University School of Medicine, 300 Pasteur Dr., Rm. G312, Stanford, CA 94305-5208, USA. bbarbara@stanford.edu
Sommer
Marvin
M
Zerboni
Leigh
L
Rajamani
Jaya
J
Arvin
Ann M
AM
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral; D006023:Glycoproteins / Q000235:genetics*; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000502:physiology; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism*; D008545:Melanoma; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D012867:Skin / Q000821:virology; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D013601:T-Lymphocytes / Q000821:virology; D015534:Trans-Activators / Q000378:metabolism*; D015533:Transcriptional Activation; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000235:genetics*
D006023:Glycoproteins; C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; D016329:Sp1 Transcription Factor; D015534:Trans-Activators; D014759:Viral Envelope Proteins