title: Binding of host-cell factors to DNA sequences in the long terminal repeat of human T-cell leukemia virus type I: implications for viral gene expression.
abstract: Efficient expression of human T-cell leukemia virus type I (HTLV-I) genes requires both host and viral proteins and is dependent on DNA sequences in the proviral long terminal repeats (LTRs). We have used DNase I-protection assays (footprinting) to construct a map of protein-DNA interactions over a 250-nucleotide region of the LTR upstream of the start site for viral RNA synthesis. We find that a host factor (host expression factor 1, or HEF-1) binds to the imperfect 21-nucleotide repeats that have previously been implicated in HTLV-I gene expression. HEF-1 binding activity is present in preparations from both lymphoid and nonlymphoid cell lines. However, the boundaries of the protected regions and the presence of a flanking DNase-hypersensitive site vary with cell type. Several regions of binding are detected in addition to the HEF-1 sites, including a complex group of sites 40-90 nucleotides upstream of the RNA start site. A comparison of HTLV-I-transformed T lymphocytes that do and do not express the viral trans-activating protein p40xI shows that none of the observed features of the DNase I footprint pattern correlate directly with the presence of this protein in the extract. These results suggest (i) that the primary recognition of promoter elements in the HTLV-I LTR involves specific interactions with host-cell proteins and (ii) that p40xI influences the activity of one or more of these proteins, rather than interacting directly with the DNA.
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
authors.forename: J K
authors.initials: JK
authors.lastname: Nyborg
authors.forename: W S
authors.initials: WS
authors.lastname: Dynan
authors.forename: I S
authors.initials: IS
authors.lastname: Chen
authors.forename: W
authors.initials: W
authors.lastname: Wachsman
mesh_terms: D001665:Binding Sites; D002460:Cell Line; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology*; D017977:Deltaretrovirus / Q000235:genetics*; D003850:Deoxyribonuclease I; D005786:Gene Expression Regulation; D006801:Humans; D012091:Repetitive Sequences, Nucleic Acid; D014158:Transcription, Genetic
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; D003850:Deoxyribonuclease I