Nuclear localization of the 82-kDa form of human choline acetyltransferase.
Choline acetyltransferase is the enzyme catalyzing synthesis of the neurotransmitter acetylcholine in cholinergic neurons. In human, transcripts encoding two forms of the enzyme with apparent molecular masses of 69 and 82 kDa are found in brain and spinal cord; the 82-kDa form differs from the 69-kDa enzyme only in terms of a 118-amino acid extension on its amino terminus. Using green fluorescent protein-tagged choline acetyltransferase, we show that the 82-kDa enzyme is targeted to nuclei of cells, whereas the 69-kDa protein is found in cytoplasm. Expression of site-directed and deletion mutants of the 82-kDa isoform reveals that the extended amino terminus contains a nuclear localization signal in the first nine amino acids which targets the protein to nucleus. This represents the first report of a neurotransmitter-synthesizing enzyme that is localized to the cell nucleus.
Resendes
M C
MC
Department of Physiology, The University of Western Ontario and the Neurodegenerative Diseases Group, The John P. Robarts Research Institute, London, Ontario N6A 5C1, Canada.
Dobransky
T
T
Ferguson
S S
SS
Rylett
R J
RJ
D000595:Amino Acid Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology*; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014162:Transfection
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D002795:Choline O-Acetyltransferase