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M9630644.TXT
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1996-02-27
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Document 0644
DOCN M9630644
TI Role of the TATA box in transcription of the mouse mammary tumor virus
long terminal repeat.
DT 9603
AU Kusk P; Carlson KE; Warren BS; Hager GL; Laboratory of Molecular
Virology, National Cancer Institute,; National Institutes of Health,
Bethesda, Maryland 20892, USA.
SO Mol Endocrinol. 1995 Sep;9(9):1180-92. Unique Identifier : AIDSLINE
MED/96064300
AB An in vitro transcription system from mammary cells was established to
study transcription of the long terminal repeat (LTR) of the mouse
mammary tumor virus (MMTV). Experiments with progressive 5'-deletion
constructs of the MMTV LTR revealed that a 19-base pair (bp) region from
-41 to -23 bp, encompassing the TATA box and flanking DNA sequence, was
as transcriptionally active as larger promoter constructs, both in
nuclear extracts from human mammary cell lines (T47D and MCF7) and a
nonmammary cell line (HeLa). The cell-free system was capable of
supporting transcriptional induction by factors binding upstream of the
TATA box, however, since purified glucocorticoid receptor-induced
transcription in larger promoter constructs encompassing the MMTV
hormone-responsive elements. Transcription from two other promoters, the
adenovirus major late promoter and the human immunodeficiency virus LTR,
also revealed a significant transcriptional contribution of upstream
elements. The 19-bp TATA region from the MMTV LTR was shown to have
considerably more activity in this transcription system than comparable
TATA regions from other promoters. Sequences critical to the MMTV TATA
region were evaluated by single base pair mutagenesis and found to
comprise a consensus TATA box sequence, TATAAAA, as well as a single A
just upstream of the TATAAAA sequence. Thus, the high level of basal
transcription observed with the TATA region from MMTV is due to a
perfect consensus TATA box sequence and a single base immediately 5'
adjacent. It is likely that the high basal rate of transcription
observed with this TATA box region on histone-free templates represents
an inappropriate level of basal expression and that a complete
evaluation of transactivation mechanisms in this system will require the
recapitulation in vitro of the chromatin-mediated repressive state that
exists in vivo.
DE Base Sequence Binding Sites Breast Neoplasms Comparative Study DNA,
Viral/*CHEMISTRY Hela Cells Human Mammary Tumor Viruses,
Mouse/*GENETICS Molecular Sequence Data Nuclear Proteins Promoter
Regions (Genetics) *Repetitive Sequences, Nucleic Acid *Transcription,
Genetic Tumor Cells, Cultured *TATA Box JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).