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PMID: 2911466 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

5' flanking and first intron sequences of the human beta-actin gene required for efficient promoter activity.

Nucleic acids research ·Vol. 17 ·No. 1 ·1989-01-11 ·Pages 253-70

Frederickson RM, Micheau MR, Iwamoto A, Miyamoto NG

Abstract

We have identified a CCAAT box element that is required for the efficient transcription of the human beta-actin gene. Both in vivo transient transfection assays in cultured HeLa cells and in vitro run-off transcription assays in HeLa whole cell extracts demonstrated the requirement of this element for efficient promoter activity. A gel mobility shift assay revealed a Hela nuclear factor that specifically interacted with the beta-actin CCAAT element in vitro; mutation of the first three base pairs of the CCAAT pentanucleotide abolished binding of this factor. Competition gel shift experiments revealed that three sequence elements located within the beta-actin promoter, each containing a CC(A/T)6GG motif similar to that contained within the c-fos serum response element, were able to bind a different nuclear factor, serum response factor (SRF). One of these CC(A/T)6GG motifs is contained within a first intron fragment that enhanced transcription from a heterologous promoter in vivo.

MeSH Terms
Actins/genetics Base Sequence DNA-Binding Proteins/metabolism Genes HeLa Cells/metabolism Humans Introns Molecular Sequence Data Mutation Promoter Regions, Genetic Restriction Mapping Transcription, Genetic Transfection
Chemicals
Actins DNA-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Frederickson R M
Division of Biological Research, Ontario Cancer Institut, Toronto, Canada.
Micheau M R
Iwamoto A
Miyamoto N G
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.1.253
Published
1989-01-11
Pages
253-70
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331549
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.1.253 PMC Full Text View journal details
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