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Oncogene. 1993 Sep;8(9):2391-7.

A trans-activator Tax of human T-cell leukemia virus type 1 binds to NF-kappa B p50 and serum response factor (SRF) and associates with enhancer DNAs of the NF-kappa B site and CArG box.

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1
Department of Cellular and Molecular Biology, University of Tokyo, Japan.

Abstract

A transcriptional activator of human T-cell leukemia virus type 1 (HTLV-1) activates at least three distinct enhancers: the viral 21-bp enhancer, the NF-kappa B binding site of the IL-2R alpha gene and the CArG box of the c-fos gene. To understand the mechanisms of Tax transactivations of the NF-kappa B enhancer and CArG box, the interactions of Tax protein with their binding factors were analysed. Using a DNA affinity precipitation (DNAP) assay, we found here that Tax associates with the DNA sequences of the NF-kappa B site and CArG box. These Tax associations with enhancers were observed only in the presence of a nuclear factor(s) and were equal to the activating capacities of Tax mutants. To identify the nuclear factor(s), we defined conditions under which no Tax binding to the NF-kappa B binding site and CArG box was detected with a nuclear extract of 293T cells. Under these conditions, transfections with cDNAs of the NF-kappa B p50 and serum response factor (SRF) produced a factor(s) that mediated Tax binding to the NF-kappa B site and the CArG box respectively. Furthermore, purified Tax protein interacted with purified NF-kappa B p50 and purified SRF, indicating their direct bindings. These observations indicate that Tax protein associates with enhancer sequences of the NF-kappa B site and CArG box through NF-kappa B p50 and SRF respectively. Previously we demonstrated that Tax interacts with CREB and CREM proteins that bind to the 21-bp enhancer DNA. These results together suggest that indirect binding of Tax to DNA through each enhancer binding protein is a general mechanism for Tax transactivation of transcription.

PMID:
8361755
[Indexed for MEDLINE]

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