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Human T-Cell Leukemia Virus Type I Tax Transactivates the Promoter of Human Prointerleukin-1 Gene Through Association With Two Transcription Factors, Nuclear Factor-Interleukin-6 and Spi-1
Junichi Tsukada,
Masahiro Misago,
Yoko Serino,
Ryosuke Ogawa,
Syuichi Murakami,
Masatsugu Nakanishi,
Shinichi Tonai,
Yoshihiko Kominato,
Isao Morimoto,
Philip E. Auron, and
Sumiya Eto
From the First Department of Internal Medicine, School of Medicine and the School of Health Sciences, University of Occupational and Environmental Health, Kitakyushu; Department of Legal Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan; and The Center for Blood Research, Harvard Medical School, Boston, MA.
The human T-cell leukemia virus type I (HTLV-I), which infects a wide variety of mammalian cells including monocytes and macrophages, encodes a transactivating protein designated as Tax. We now report that Tax induces the human prointerleukin-1 (IL1B) gene promoter in monocytic cells. In our transient transfection assays using human THP-1 monocytic cells, a chloramphenicol acetyltransferase (CAT) construct containing the IL1B promoter sequence between positions -131 and +12 showed an approximately 90-fold increase in activity following cotransfection of a Tax expression vector. Moreover, Tax synergized with lipopolysaccharide (LPS) to induce the IL1B promoter activity. Analyses of specific nucleotide substitutions further indicated that the Tax-induced transcriptional activation requires two transcription factor binding motifs within the IL1B promoter; one is a binding site for nuclear factor (NF)-IL6 (CCAAT/enhancer binding protein , C/EBP), which belongs to the basic region-leucine zipper (bZIP) family and the other for Spi-1 (PU.1), which is an Ets family protein found principally in monocytes, macrophages, and B lymphocytes. In electrophoretic mobility shift assays (EMSA) using in vivo THP-1 nuclear extracts, Tax expression in THP-1 monocytic cells significantly increased binding of the two factors to their target IL1B promoter sequences. However, in contrast to NF-IL6 and Spi-1, DNA binding activity of Oct-1, an ubiquitously expressed octamer-binding protein was not affected by Tax. Additional EMSA using in vitro translated proteins also showed that recombinant Tax enhances DNA binding of both of recombinant NF-IL6 and Spi-1 proteins. These data were supported by our glutathione S-transferase (GST)-pulldown data, which indicated that Tax physically interacts with the two proteins. Based on the results obtained from the present study, we conclude that the IL1B promoter is a Tax-responsive sequence as a result of ability of Tax to induce binding of NF-IL6 and Spi-1 to the IL1B promoter sequence through protein-protein interaction.
Blood, Vol. 90 No. 8 (July 15), 1997:
pp. 3142-3153
© 1997 by The American Society of Hematology.
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