Gene expression of mouse choline acetyltransferase: Alternative splicing and identification of a highly active promoter region

Hidemi Misawa, Kayoko Ishii, Takeo Deguchi

Research output: Contribution to journalArticle

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Abstract

Seven types of mRNA for choline acetyltransferase that differ in the 5′-noncoding region were identified in the mouse spinal cord by cDNA cloning and polymerase chain reaction. Among these transcripts, the M-type mRNA corresponding to the previously cloned mouse cDNA was most abundant in the spinal cord of mouse. A mouse genomic DNA clone containing the 5′-region of choline acetyltransferase mRNA was isolated and sequenced. Comparison of the sequences between the cDNAs and the genomic DNA revealed that the different mRNA species were transcribed from different promoter regions and produced by differential splicing. Two murine cholinergic cell lines, NS20Y and NG108-15, were shown to express the M-type mRNA almost exclusively, and were therefore used to study transcription of M-type mRNA. Fragments of the 5′-region of choline acetyltransferase gene were ligated with chloramphenicol acetyltransferase reporter gene and introduced into cultured cells. The fragment from -2752 to +46, which contained the M-type exon, a TATA-box like element upstream of the M-type exon, and the downstream intron, induced a significant expression of CAT activity in neuronal but not in non-neuronal cell lines. This result indicates that this region of choline acetyltransferase gene contains elements that regulate neuron-specific expression of choline acetyltransferase activity. However, there was no parallel correlation between reporter gene expression in the transfected cells and intrinsic choline acetyltransferase activity in these neuronal cell lines. Possible mechanisms that would explain this observation are discussed.

Original languageEnglish
Pages (from-to)20392-20399
Number of pages8
JournalJournal of Biological Chemistry
Volume267
Issue number28
Publication statusPublished - 1992 Oct 5
Externally publishedYes

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Choline O-Acetyltransferase
Alternative Splicing
Genetic Promoter Regions
Gene expression
Gene Expression
Messenger RNA
Cells
Complementary DNA
Genes
Reporter Genes
Cell Line
Exons
Spinal Cord
TATA Box
Chloramphenicol O-Acetyltransferase
Cloning
Polymerase chain reaction
DNA
Transcription
Introns

ASJC Scopus subject areas

  • Biochemistry

Cite this

Gene expression of mouse choline acetyltransferase : Alternative splicing and identification of a highly active promoter region. / Misawa, Hidemi; Ishii, Kayoko; Deguchi, Takeo.

In: Journal of Biological Chemistry, Vol. 267, No. 28, 05.10.1992, p. 20392-20399.

Research output: Contribution to journalArticle

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