JBC INTERFERin siRNA transfection reagent

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Volume 272, Number 8, Issue of February 21, 1997 pp. 5208-5213
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Nuclear ADP-ribosylation Factor (ARF)- and Oleate-dependent Phospholipase D (PLD) in Rat Liver Cells
INCREASES OF ARF-DEPENDENT PLD ACTIVITY IN REGENERATING LIVER CELLS

(Received for publication, August 29, 1996, and in revised form, November 5, 1996)

Yoshiko Banno Dagger , Keiko Tamiya-Koizumi , Hideko Oshima , Akemi Morikawa Dagger , Shonen Yoshida and Yoshinori Nozawa Dagger

From the Dagger  Department of Biochemistry, Gifu University School of Medicine, Tsukasamachi-40, Gifu 500 and the  Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-Ku, Nagoya, Aich 466, Japan

Two forms of phospholipase D (PLD) have been found to be present in nuclei isolated from rat hepatocytes by measuring phosphatidylbutanol produced from exogenous radiolabeled phosphatidylcholine in the presence of butanol. In nuclear lysates from either rat liver or ascites hepatoma AH 7974 cells, the PLD activity was markedly stimulated by a recombinant ADP-ribosylation factor (rARF) in the presence of the guanosine 5'-O-(3-thiotriphosphate) (GTPgamma S) and phosphatidylinositol 4,5-bisphosphate. ATP and phorbol-12-myristate 13-acetate had no synergistic effect on this PLD activity. On the other hand, the nuclear PLD was stimulated by unsaturated fatty acids, especially by oleic acid. The ARF-dependent nuclear PLD activity was increased in the S-phase of the regenerating rat liver after partial hepatectomy and also was much higher in AH 7974 cells than in the resting rat liver. In contrast, the levels of the oleate-dependent PLD activity remained constant throughout the cell cycle in liver regeneration. The intranuclear levels of the stimulating proteins of the nuclear PLD activity, e.g. ARF, RhoA, and protein kinase Cdelta increased in the S-phase of the regenerating liver. These results suggested that the nuclear ARF-dependent PLD activity may be associated with cell proliferation.


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