TY - JOUR
T1 - Human JC virus nuclear factor 1 binding motifs and large tumor antigen region required for transactivation of late promoter
AU - Kumar, Kotlo U.
AU - Devireddy, Laxminarayana R.
AU - Tang, Shou Ching
AU - Pater, Alan
AU - Pater, Mary M.
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1996/8
Y1 - 1996/8
N2 - 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 JCV(L) 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 II/III 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 JCV(L), 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 JCV(L) 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.
AB - 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 JCV(L) 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 II/III 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 JCV(L), 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 JCV(L) 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.
KW - JC virus
KW - Late promoter
KW - Nuclear factor 1 sites
KW - T-Ag
KW - Transactivation
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U2 - 10.1046/j.1471-4159.1996.67020473.x
DO - 10.1046/j.1471-4159.1996.67020473.x
M3 - Article
C2 - 8764570
AN - SCOPUS:0030055607
SN - 0022-3042
VL - 67
SP - 473
EP - 481
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
IS - 2
ER -