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PMID: 2355914 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Muscle-specific activity of the skeletal troponin I promoter requires interaction between upstream regulatory sequences and elements contained within the first transcribed exon.

Molecular and cellular biology ·Vol. 10 ·No. 7 ·1990-07-00 ·Pages 3468-82

Nikovits W, Mar JH, Ordahl CP

Abstract

Expression of the skeletal troponin I (sTnI) gene is regulated transcriptionally in a muscle-specific fashion. We show here that the region of the sTnI gene between -160 and +61 (relative to the transcription initiation site) is able to direct expression of the bacterial chloramphenicol acetyltransferase (CAT) gene is muscle cultures at a level approximately 100 times higher than in fibroblast cultures. RNA analysis demonstrated that transcription of the CAT gene was initiated at the same site as transcription of the endogenous sTnI gene and that CAT activity levels were approximately proportional to CAT mRNA levels. Deletion analysis demonstrated that the region between nucleotides -160 and -40 contained sequences essential for full promoter activity. Surprisingly, 3' deletion analysis indicated that the first exon (-6 to +61) of the sTnI gene was also required for full activity of the sTnI promoter in skeletal muscle cells. Chimeric promoter experiments, in which segments of the sTnI and the herpes simplex virus thymidine kinase promoter were interchanged, indicated that reconstitution of a muscle-specific promoter required inclusion of both the upstream and exon I regions of the sTnI gene. Exon I, and the region immediately upstream, showed DNase protection over sequence motifs related to those found in other genes, including the tar region of human immunodeficiency virus type 1. These results demonstrate that expression of the sTnI promoter in embryonic skeletal muscle cells requires complex interaction between two separate promoter regions, one of which resides within the first 61 transcribed nucleotides of the gene.

MeSH Terms
Animals Base Sequence Cell Nucleus/metabolism Chick Embryo Chromosome Deletion Cloning, Molecular Exons Gene Expression Regulation Genes Genes, Regulator Molecular Sequence Data Muscles/metabolism Plasmids Promoter Regions, Genetic Restriction Mapping Transcription, Genetic Troponin/genetics Troponin I
Chemicals
Troponin Troponin I
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nikovits W
Department of Anatomy, University of California, San Francisco 94143.
Mar J H
Ordahl C P
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.7.3468-3482.1990
Published
1990-07-00
Pages
3468-82
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360782
Subset
IM
Grants
NIGMS NIH HHS · GM32018 · United States
NHLBI NIH HHS · HL35561 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.7.3468-3482.1990 PMC Full Text View journal details
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