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Transcriptional regulation of the human interleukin-6 gene promoter in
human T-cell leukemia virus type I-infected T-cell lines: evidence for the
involvement of NF-kappa B
N Mori, F Shirakawa, H Shimizu, S Murakami, S Oda, K Yamamoto and S Eto
First Department of Internal Medicine, School of Medicine, University of
Occupational and Environmental Health, Kitakyushu, Japan.
Freshly isolated leukemic cells from patients with adult T-cell leukemia
(ATL) and human T-cell leukemia virus type I (HTLV-I)-infected T-cell lines
constitutively produce high levels of interleukin-6 (IL-6) protein and
mRNA. To clarify the mechanisms that lead to the activation of IL-6 gene in
HTLV-I-infected cells, we first studied the regulatory regions in the IL-6
gene transcription by transfection of chloramphenicol acetyltransferase
(CAT) reporter plasmids containing the IL-6 promoter. When transfected into
HTLV-I-infected T-cell lines MT-2 and HUT-102, IL-6 promoter/CAT plasmids
were strongly activated without any stimulation. By deletion analysis of 5'
upstream region of IL-6 promoter, the DNA region between -73 and -59 bp
from the transcription start site of IL-6 gene was important in the
expression of IL-6/CAT activities in HTLV-I-infected cells. This region
contains nuclear factor (NF)-kappa B binding site. The site-directed
mutation of the kappa B motif in IL-6/CAT plasmid resulted in the complete
abrogation of IL-6 promoter activity in these cells. Furthermore, when IL-6
promoter/CAT plasmid was introduced into an HTLV-I-uninfected T- cell line,
Jurkat, IL-6 promoter activity was silent in the basal level, but strongly
increased by the cotransfection with an HTLV-I tax expression plasmid.
However, tax expression plasmid showed no transactivation activity, when
kappa B site was mutated in IL-6 promoter/CAT plasmid. We found that the
IL-6 kappa B site specifically formed a complex with NF-kappa B-containing
nuclear extracts from MT-2 and HUT-102 cells. Finally, transfection of
HTLV-I tax into Jurkat cells resulted in induction of specific binding of
nuclear extracts to the NF-kappa B sequence. These results strongly suggest
that HTLV-I tax gene may transactivate IL-6 gene through kappa B site in
HTLV-I- positive T-cell lines and activation of NF-kappa B may be crucial
in HTLV-I-induced IL-6 gene activation in ATL.
Volume 84,
Issue 9,
pp. 2904-2911,
11/01/1994
Copyright © 1994 by The American Society of Hematology
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