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PMID: 2950379 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Specific cellular proteins bind to critical promoter sequences of the adenovirus early EIIa promoter.

Nucleic acids research ·Vol. 15 ·No. 2 ·1987-01-26 ·Pages 509-27

Boeuf H, Zajchowski DA, Tamura T, Hauss C, Kédinger C

Abstract

As an approach to the identification of essential factors required for specific expression of the adenovirus type 2 EIIaE early (EIIaE) promoter, an in vitro system was established. Under appropriate conditions, using crude extracts of non-infected HeLa cells, efficient and accurate EIIaE expression has been reproduced. As in vivo, this transcription was strongly dependent upon the integrity of two non-consensus TATA-like elements, T1 and T2, corresponding to the major (EIIaE1) and minor (EIIaE2) start sites, respectively, as well as upon intact upstream elements (A, between -40 and -50 and B, between -70 and -90) common to both overlapping promoters. The implication of specific DNA-binding proteins in the transcriptional effects mediated by these elements was demonstrated by DNAse I footprinting analyses. Both crude nuclear extracts and partially purified fractions confer specific protection against DNAse I digestion to the T1 and B promoter elements defined above, and to a far upstream region (element C, between -110 and -150), which has previously been identified as a weaker promoter element by in vivo transcriptional studies. Separation of the T1 recognition factor from those which bind to the upstream elements B and C by chromatographic fractionation of the extracts has also been achieved.

MeSH Terms
Adenovirus Early Proteins Adenoviruses, Human/genetics,metabolism Antigens, Viral, Tumor/genetics Base Sequence DNA, Viral/genetics,metabolism Genes Genes, Viral HeLa Cells/metabolism Humans Nucleoproteins/isolation & purification,metabolism Oncogene Proteins, Viral/genetics Promoter Regions, Genetic Protein Binding Transcription, Genetic
Chemicals
Adenovirus Early Proteins Antigens, Viral, Tumor DNA, Viral Nucleoproteins Oncogene Proteins, Viral
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Boeuf H
Zajchowski D A
Tamura T
Hauss C
Kédinger C
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.2.509
Published
1987-01-26
Pages
509-27
Language
English
Region
England
NLM ID
0411011
PMCID
PMC340449
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/15.2.509 PMC Full Text View journal details
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