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J Biol Chem, Vol. 274, Issue 30, 21078-21084, July 23, 1999
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From the 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 Department of Pharmacology and
§ Department of Biochemistry and Molecular Biology, Mayo
Clinic, Rochester, Minnesota 55905
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.
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