Figure 1
From: SF2/ASF binding region within JC virus NCCR limits early gene transcription in glial cells
The “CR3” region within JCV NCCR is the target for SF2/ASF. A. Sequence alignment for the NCCR region of JCV. CLUSTAL sequence alignment was performed for JCV Mad1 and Archetype strains (gene bank accession number NC_001699). The positions of replication origin (ORI), CR1, CR2, CR3, CR4 and 98-bp-tandem repeats are illustrated in Mad1 strain. CR3 region is highlighted with sequences in red. Numbering is relative to the Mad-1 strain of JCV. B. Upper panel: Schematic presentation of Mad1-WT, Mad1-(1X98), and ΔCR3-(1X73) promoter. Arrows point the position of forward (PF) and reverse (PR) primers used in ChIP experiments. Lower panel: PHFA cells were transiently transfected with pCGT7-SF2/ASF expression vector (SF2) or pCGT7 vector alone (vector) and pBLCAT3-JCV-RR-WT, pBLCAT3-JCV-RR-(1X98), and pBLCAT3-JCV-RR-ΔCR3-(1X73) reporter plasmids. Cells were cross-linked and ChiP assay was performed using antibody to T7-tagged SF2/ASF (lanes 4 to 12). In lanes 1, 2 and 3, pBLCAT3-JCV-RR-WT, pBLCAT3-JCV-RR-(1X98), and pBLCAT3-JCV-RR-ΔCR3-(1X73) reporter plasmid DNAs were used as positive controls. C. Western blot analyzes of whole cell lysates from panel A (lanes 4 to 12), using specific antibodies against SF2/ASF and Tubulin.