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PMID: 3785201 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Delimitation and characterization of cis-acting DNA sequences required for the regulated expression and transcriptional control of the chicken skeletal alpha-actin gene.

Molecular and cellular biology ·Vol. 6 ·No. 7 ·1986-07-00 ·Pages 2462-75

Bergsma DJ, Grichnik JM, Gossett LM, Schwartz RJ

Abstract

We have previously observed that DNA sequences within the 5'-flanking region of the chicken skeletal alpha-actin gene harbor a cis-acting regulatory element that influences cell type and developmental stage-specific expression (J. M. Grichnik, D. J. Bergsma, and R. J. Schwartz, Nucleic Acids Res 14:1683-1701, 1986). In this report we have constructed unidirectional 5'-deletion and region-specific deletion-insertion mutations of the chicken skeletal alpha-actin upstream region and inserted these into the chloramphenicol acetyltransferase expression vector pSV0CAT. These constructions were used to locate DNA sequences that are required for developmental modulation of expression when transfected into differentiating myoblasts. With this assay we have delimited the 5' boundary of a cis-acting regulatory element to ca. 200 base pairs upstream of the mRNA cap site. In addition, we have preliminarily identified DNA sequences that may be important subcomponents within this element. A second major focus of this study was to identify those DNA signals within the regulatory element that control transcription. Toward this end, the expression phenotypes of progressive 5'-deletion and deletion-insertion mutants of the 5'-flanking region of the chicken skeletal alpha-actin gene were assayed in microinjected Xenopus laevis oocytes. These experiments defined a cis-acting transcriptional control region having a 5' border 107 base pairs preceding the alpha-actin RNA cap site. Proximal and distal functionally important regions of DNA were identified within this element. These DNA signals included within their DNA sequences the "CCAAT" and "TATA" box homologies.

MeSH Terms
Actins/genetics Animals Base Sequence Chickens Chimera Chromosome Deletion DNA/analysis Female Gene Expression Regulation Microinjections Muscles/analysis Mutation Oocytes/analysis Plasmids Transcription, Genetic Xenopus
Chemicals
Actins DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bergsma D J
Grichnik J M
Gossett L M
Schwartz R J
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53 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.7.2462-2475.1986
Published
1986-07-00
Pages
2462-75
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367800
Subset
IM
Grants
NINDS NIH HHS · NS-15050 · United States
Databases
GENBANK
J03367, M13631
External Links
PubMed source DOI: 10.1128/mcb.6.7.2462-2475.1986 PMC Full Text View journal details
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