       Document 0629
 DOCN  M94A0629
 TI    Multiple proteins interact over the USF binding site in the human
       immunodeficiency virus type 1 long terminal repeat.
 DT    9412
 AU    Tanskanen E; Deacon N; Churchill M; Doherty R; Macfarlane Burnet Centre
       for Medical Research, Fairfield, Vic.
 SO    Annu Conf Australas Soc HIV Med. 1993 Oct 28-30;5:69 (abstract no. FB4).
       Unique Identifier : AIDSLINE ASHM5/94349026
 AB    HIV-1 gene expression and viral replication are regulated by several
       viral and cellular proteins, most commonly through binding sites within
       the long terminal repeats (LTRs) of the viral genome. We have examined
       binding of transcription factors over the region from nt -141 to nt -180
       relative to transcription initiation using a novel approach in which a
       series of five oligonucleotides were constructed with 13bp overlapping
       altered sequences as compared to the wild-type HXB2 sequence. These
       oligonucleotides were used in excess as competitors for protein binding
       in electrophoretic mobility shift assays. Three of the five
       oligonucleotides lost the ability to compete for proteins binding to the
       wildtype sequence, localising nuclear factor binding to sequences
       upstream of nt -159. This region contains the motif CACATG which has
       previously been shown by DNAse footprinting to bind the transcription
       factor USF (Giacca et al, Virology 1992; 186:133-147), however our data
       indicate a more precise region of interaction. Examination of binding
       from nuclear extracts of the lymphoblastoid cell lines CEM and MT-2 as
       well as the epithelial HeLa cell line has shown that the interaction
       involves more than one protein and that the different cell lines exhibit
       different binding patterns over this region of the LTR. We have
       calculated the approximate size of these DNA-protein complexes by UV
       crosslinking and south-western analyses.
 DE    Cell Line  DNA-Binding Proteins/*GENETICS  Gene Expression Regulation,
       Viral/PHYSIOLOGY  Human  HIV-1/*GENETICS  Repetitive Sequences, Nucleic
       Acid/*GENETICS  Transcription Factors/*GENETICS  Virus
       Replication/*GENETICS  MEETING ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

