Novel messenger RNA and alternative promoter for murine acetylcholinesterase

Elena Atanasova, Sharon Chiappa, Eric D Wieben, William Stephen Brimijoin

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A portion of the 5'-flanking region of murine acetylcholinesterase was cloned from genomic DNA by 5'-rapid amplification of genomic ends, identified in a mouse genomic library, and sequenced. Multiple potential binding sites for universal and tissue-specific transcription factors were suggestive of a promoter region within this DNA sequence. Potential promoter activity was confirmed by coupling the new sequence to the open reading frame of a luciferase reporter gene in transient expression experiments with nerve and muscle cells. 5'-Rapid amplification of cDNA ends with templates from multiple sources revealed a novel transcription start site (at position - 626, relative to translation start), located 32 bases downstream from a TATAA sequence. This start site appeared to mark a novel exon (1a) comprising 291 base pairs between positions -335 and -626, relative to the translation start. Supporting this conclusion, polymerase chain reactions with cDNA from mouse brain, heart, and other tissues, consistently amplified a transcript containing the exon 1a sequence fused to the invariant sequence beginning at position -22 in exon 2, but lacking exon 1. Northern blot analyses confirmed the in vivo expression of exon 1a-containing transcripts, especially in heart, brain, liver, and kidney. These results indicate that the murine acetylcholinesterase gene has a functioning alternative promoter that may influence expression of acetylcholinesterase in certain tissues.

Original languageEnglish (US)
Pages (from-to)21078-21084
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number30
DOIs
StatePublished - Jul 23 1999

Fingerprint

Acetylcholinesterase
Exons
Messenger RNA
Tissue
Amplification
Brain
Complementary DNA
Genes
Genomic Library
5' Flanking Region
Transcription Initiation Site
Polymerase chain reaction
DNA sequences
Luciferases
Reporter Genes
Genetic Promoter Regions
Base Pairing
Northern Blotting
Liver
Muscle Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Novel messenger RNA and alternative promoter for murine acetylcholinesterase. / Atanasova, Elena; Chiappa, Sharon; Wieben, Eric D; Brimijoin, William Stephen.

In: Journal of Biological Chemistry, Vol. 274, No. 30, 23.07.1999, p. 21078-21084.

Research output: Contribution to journalArticle

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