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PMID: 2826127 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro binding of cell-specific and ubiquitous nuclear proteins to the octamer motif of the SV40 enhancer and related motifs present in other promoters and enhancers.

The EMBO journal ·Vol. 6 ·No. 10 ·1987-10-00 ·Pages 3015-25

Rosales R, Vigneron M, Macchi M, Davidson I, Xiao JH, Chambon P

Abstract

We have used the gel retardation and DNase I footprinting assays to investigate the in vitro binding of nuclear proteins to the octamer motif present in domain A of the SV40 enhancer and in other enhancer and promoter elements. Three apparently cell-specific (oct-B1A, oct-B1B and oct-B2) and one ubiquitous (oct-B3) proteins were detected in various lymphoid and non-lymphoid cell extracts. We show that the previously described 'ubiquitous' NF-A1 factor may correspond in fact to two proteins, oct-B1A in HeLa cells and oct-B1B in lymphoid cells. Interestingly, the HeLa cell protein oct-B1A formed a complex with the SV40 octamer, which could be detected in gel retardation, but not in DNase I footprinting assays. This absence of protection from DNase I digestion correlates with the inactivity of the SV40 octamer in HeLa cells in vivo. We have also found that the in vitro interaction between the SV40 octamer motif and the lymphoid cell-specific protein oct-B2 was negatively modulated by a component present in the nuclear extracts from several lymphoid cell lines. The interactions between the multiple octamer-binding proteins and the related octamer motifs present in other promoter and enhancer elements were systematically compared and the possible role of these proteins in the control of transcription is discussed.

MeSH Terms
Base Sequence Cell Line Enhancer Elements, Genetic Genes, Viral HeLa Cells/metabolism Humans Kinetics Molecular Sequence Data Nuclear Proteins/metabolism Oligodeoxyribonucleotides Protein Binding Simian virus 40/genetics
Chemicals
Nuclear Proteins Oligodeoxyribonucleotides
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Rosales R
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique de de l'INSERM, Faculté de Médicine, Strasbourg, France.
Vigneron M
Macchi M
Davidson I
Xiao J H
Chambon P
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1987-10-00
Pages
3015-25
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553738
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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