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J. Virol., Jan 1997, 337-344, Vol 71, No. 1
Copyright © 1997, American Society for Microbiology

Function of the human T-cell leukemia virus type 1 21-base-pair repeats in basal transcription

MK Barnhart, LM Connor and SJ Marriott
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030, USA.

The human T-cell leukemia virus type 1 (HTLV-1) promoter contains three copies of an imperfect 21-bp repeat called Tax-responsive element (TRE1). To examine the role of individual TRE1 sequences in basal transcription of the HTLV-1 promoter, site-directed mutations were generated in all possible combinations of one, two, or all three TRE1 elements in the viral long terminal repeat (LTR) and tested in vivo for transcriptional activity. Mutation of the middle TRE1 resulted in the greatest reduction in basal activity. Electrophoretic mobility shift analysis demonstrated that the protein complexes bound to each of the three TRE1 sequences were not identical. The complexes formed with the TATA-distal and middle TRE1s were dependent on the core cyclic AMP response element (CRE) found in all three TRE1s, while the cellular transcription factor Sp1 bound the TATA-proximal TRE1 in a CRE- independent manner. Sp1 binding produced a footprint on the viral LTR which covered the 5' region of the proximal TRE1. Mixing experiments demonstrated that the bindings of CREB and Sp1 to the proximal TRE1 were mutually exclusive. Sp1 was able to activate transcription both from the complete LTR and from the proximal TRE1 alone. These studies demonstrate that the TRE1 elements in the HTLV-1 LTR are functionally nonequivalent and suggest that Sp1 can influence HTLV-1 basal transcription.

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