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PMID: 2493627 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Point mutational analysis of the human c-fos serum response factor binding site.

Nucleic acids research ·Vol. 17 ·No. 3 ·1989-02-11 ·Pages 1177-95

Leung S, Miyamoto NG

Abstract

A series of point mutants were generated in the human c-fos dyad symmetry element (DSE), found within the c-fos serum response element, to study the sequence requirements for its interaction with the human HeLa cell serum response factor (SRF). Plasmids that contain base substitutions within a core CC(A/T)6GG motif in the center of the DSE did not compete, or competed very poorly, with the wild-type c-fos DSE for formation of a specific SRF-DSE complex in vitro. The CC(A/T)6GG motif is not sufficient for maximal binding of SRF, as several plasmids that contain base substitutions in the sequences flanking this core motif competed either poorer or better than the wild-type c-fos DSE for SRF binding. Evidence is presented that supports the idea that SRF binds in a symmetrical fashion. Results of in vivo transient expression assays in HeLa cells suggest that negative regulation of c-fos transcription observed in serum-deprived cells is mediated through SRF binding to the DSE.

MeSH Terms
Base Sequence Binding, Competitive Blood Physiological Phenomena DNA Mutational Analysis/methods DNA-Binding Proteins/genetics Electrophoresis, Polyacrylamide Gel HeLa Cells Humans Molecular Sequence Data Nuclear Proteins/genetics,metabolism Nucleic Acid Conformation Promoter Regions, Genetic Proto-Oncogene Proteins/blood,genetics Proto-Oncogene Proteins c-fos Regulatory Sequences, Nucleic Acid Serum Response Factor
Chemicals
DNA-Binding Proteins Nuclear Proteins Proto-Oncogene Proteins Proto-Oncogene Proteins c-fos Serum Response Factor
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Leung S
Division of Biological Research, Ontario Cancer Institute, Toronto, Canada.
Miyamoto N G
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.3.1177
Published
1989-02-11
Pages
1177-95
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331729
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.3.1177 PMC Full Text View journal details
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