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TR TITLE AUTHORS KEYWORDS

MATERIALS & METHODS

 MICROSCOPIC TECHNIQUES SPECIES MORPHOLOGY CELL LINE
626 Nuclear Localization of the 82-kDa Form of Human Choline
Acetyltransferase		 Manuel C. Resendes, Tomas Dobransky, Stephen S. G. Ferguson, R. Jane Rylett Choline acetyltransferase, enzyme catalyzing
synthesis, brain, spinal cord, amino acid, cell nucleus

For confocal microscopy, cells were replated at 24 h after transfection
onto 35-mm dishes containing a 1-cm glass-bottomed center well (Plastek Cultureware, Ashland, MA) in growth medium supplemented with 20 mM HEPES buffer, pH 7.4.

 confocal microscopy HEK-293
  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.  

 

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