Intracellular compartmentation of diadenosine tetraphosphate (Ap4A) and dTTP in rat liver

Abstract

1. The intracellular compartmentation of diadenosine tetraphosphate (Ap4A) and of dTTP was studied in rat liver cells using non-aqueous glycerol for the isolation of cell nuclei. 2. This method allows a stepwise removal of cytoplasm from the nuclei. 3. The decrease in Ap4A or dTTP during the process was compared to the simultaneous decrease in RNA, which was taken to represent the cytoplasm. 4. In regenerating liver excised 24 hr after partial hepatectomy, Ap4A was almost equally distributed between the nucleus and cytoplasm. 5. In livers from unoperated control rats, the nuclear concentration of Ap4A was slightly elevated compared to that of whole cells. dTTP was only investigated in regenerating liver. 6. Significantly higher concentrations were found in the nuclear fractions. 7. The purest nuclei contained about 26% of whole cell levels of dTTP, while their RNA values had decreased to 7% of the whole cell RNA. 8. Considering that the liver cell nucleus comprises about 7% of the entire cell mass, a nuclear dTTP concentration of 26% indicates significantly higher dTTP levels in the nuclear compartment than in the cytoplasm of regenerating rat liver cells.