title: Human JC virus nuclear factor 1 binding motifs and large tumor antigen region rquired for transactivation of late promoter.
abstract: The nuclear factor 1 (NF-1) motifs, NF-1 II/III, In the two 98-bp repeats of the transcription-regulatory region of JC virus (JCV), have a critical role in brain-specific transcription from the JCV early promoter-enhancer. In this study, the role of these motifs in transactivation of the JCV late promoter-enhancer (JCVL) was examined in differentiating glial P19 embryonal carcinoma cells. The expression of papovaviral large tumor antigen (T-Ag) in the glial cells was shown by double immunofluorescence assays. By using site-directed mutagenesis and in vivo assays, the two wild-type NF-1 II/III sites, but not the third site, were found to be essential for the transactivation of JCVL by JCV T-Ag. In vitro transcription assays confirmed this specific transactivation and the transactivation was abolished by T-Ag antibody. In electrophoretic mobility shift assays, expression of JCV T-Ag increased the binding of a factor(s) to the 98-bp repeat. T-Ag antibody abolished the increase of binding. Binding assays with oligonucleotides of NF-1 11/111 motifs showed that the increased binding specifically required the wild-type NF-1 II/III sequences and confirmed the requirement of T-Ag. To determine the region of T-Ag necessary for transactivation Of JCVL, the coding sequences were mutated. The amino-terminal region of JCV Ag in amino acids 1-437 was essentially required for efficient transactivation. These results indicated that transactivation of JCVL and increased binding require a factor(s) found specifically in glial cells, the JCV NF-1 II/III sites, and the T-Ag amino-terminal region.
authors.affiliation: Basic Medical Sciences, Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
authors.forename: K U
authors.initials: KU
authors.lastname: Kumar
authors.forename: L R
authors.initials: LR
authors.lastname: Devireddy
authors.forename: S C
authors.initials: SC
authors.lastname: Tang
authors.forename: A
authors.initials: A
authors.lastname: Pater
authors.forename: M M
authors.initials: MM
authors.lastname: Pater
mesh_terms: D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D001921:Brain / Q000382:microbiology; D022762:CCAAT-Enhancer-Binding Proteins; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D007577:JC Virus / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D051839:NFI Transcription Factors; D009457:Neuroglia / Q000382:microbiology; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes / Q000737:chemistry; D011401:Promoter Regions, Genetic; D014157:Transcription Factors; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D051840:Y-Box-Binding Protein 1
chemical_list: D000957:Antigens, Viral, Tumor; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D051839:NFI Transcription Factors; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1; C070856:YBX1 protein, human