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Gel electrophoresis mobility shift and DNase I footprint assays detect a cellular nuclear protein in extracts made from uninfected human cells which binds to a downstream promoter sequence (DPS) in the human adenovirus 2 major late promoter. By DNase I footprint and mutation analyses, we have determined that this new regulatory element extends from positions +146 to +165 (relative to the cap site at position +1). We show by UV cross-linking that a 40-kDa polypeptide specifically binds to this region. Mutations within the DPS which decrease protein binding by 80-90% also cause a 2.5-3-fold decrease in in vitro major late promoter transcription efficiency. Alteration of the template in the 5'-flanking region of the DPS does not affect nuclear protein binding or transcription efficiency. Interestingly, a T----G transversion at position +160 which increases protein binding also impairs promoter activity.
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
June 1990,
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R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
July 1986,
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R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
January 1990,
Journal of virology,
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
December 1994,
Journal of virology,
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
August 1992,
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R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
February 1981,
Proceedings of the National Academy of Sciences of the United States of America,
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
June 1991,
Nucleic acids research,
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
May 1990,
The EMBO journal,
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
September 1992,
Journal of virology,
R B Cohen, and
L Yang, and
J A Thompson, and
B Safer
August 1993,
The Journal of biological chemistry,
