title: Two mRNAs are transcribed from the human gene for choline acetyltransferase.
abstract: The product of the choline acetyltransferase (ChAT) gene is the enzyme that synthesizes the neurotransmitter acetylcholine. A 14.4-kb portion of the human ChAT gene contains 7 exons, which are estimated to comprise approximately one-third of the human protein coding sequence by comparison with porcine ChAT mRNA. Two of the exons were used to identify polyadenylated human ChAT gene transcripts on Northern blots. An exon with 84% identity to the region of porcine ChAT mRNA that codes for the amino terminus of the corresponding protein detected 6,000- and 2,300-nucleotide mRNAs in RNA isolated from human CHP134 neuroblastoma cells. Only the 2,300-nucleotide mRNA was detected by a second probe containing an exon with 96% identity to porcine ChAT mRNA in the domain that encodes amino acids 204-263 of the predicted porcine ChAT protein. Further evidence that two species of human mRNA are produced from the human ChAT gene was obtained from nuclease protection assays using an antisense RNA probe prepared from a human ChAT cDNA clone. Total RNA isolated from either CHP134 cells or adult human nucleus basalis protected 525- and 400-nucleotide fragments of this probe, confirming the presence of two species of RNA that differ by the inclusion of an internal exon. cDNA clones of each of these transcripts have been isolated. Their sequences suggest that the 2,300-nucleotide mRNA encodes enzymatically active human ChAT, while translation of the 6,000-nucleotide mRNA would be terminated prematurely by a shift in the reading frame. These results indicate that a complex pattern of transcription produces two mRNAs with different coding potentials from the human ChAT gene.
authors.affiliation: Department of Molecular and Cellular Pharmacology, University of Miami School of Medicine, FL 33101.
authors.forename: M V
authors.initials: MV
authors.lastname: Lorenzi
authors.forename: A C
authors.initials: AC
authors.lastname: Trinidad
authors.forename: R
authors.initials: R
authors.lastname: Zhang
authors.forename: W L
authors.initials: WL
authors.lastname: Strauss
mesh_terms: D015152:Blotting, Northern; D002795:Choline O-Acetyltransferase / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D005091:Exons; D016368:Frameshift Mutation; D015870:Gene Expression; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D015347:RNA Probes; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000235:genetics*; D015183:Restriction Mapping; D017423:Sequence Analysis, RNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
chemical_list: D015342:DNA Probes; D015347:RNA Probes; D016372:RNA, Antisense; D012333:RNA, Messenger; D004247:DNA; D002795:Choline O-Acetyltransferase