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Originally published In Press as doi:10.1074/jbc.M006533200 on February 13, 2001

J. Biol. Chem., Vol. 276, Issue 21, 17754-17761, May 25, 2001
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Presence and Activation of Nuclear Phosphoinositide 3-Kinase C2beta during Compensatory Liver Growth*

Aleksandra Sindstrokic'Dagger §, Aleksandra AleksandrovaDagger §, Alan P. Fields, Stefano Volinia||, and Hrvoje Banfic'Dagger **

From the Dagger  Department of Physiology and Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Salata 3, Zagreb 10,000, Croatia, the  Sealy Center for Cancer Cell Biology, University of Texas Medical Branch, Galveston, Texas 77555-1048, and the || Dip. di Morfologia ed Embriologia, Universita degli Studi, Via Fossato di Mortara 64/b, Ferrara 44100, Italy

Highly purified liver nuclei incorporated radiolabeled phosphate into phosphatidylinositol 4-phosphate (PtdIns(4)P), PtdIns(4,5)P2, and PtdIns(3,4,5)P3. When nuclei were depleted of their membrane, no radiolabeling of PtdIns(3,4,5)P3 could be detected showing that within the intranuclear region there are no class I phosphoinositide 3-kinases (PI3K)s. In membrane-depleted nuclei harvested 20 h after partial hepatectomy, the incorporation of radiolabel into PtdIns(3)P was observed together with an increase in immunoprecipitable PI3K-C2beta activity, which is sensitive to wortmannin (10 nM) and shows strong preference for PtdIns over PtdIns(4)P as a substrate. On Western blots PI3K-C2beta revealed a single immunoreactive band of 180 kDa, whereas 20 h after partial hepatectomy gel shift of 18 kDa was noticed, suggesting that observed activation of enzyme is achieved by proteolysis. When intact membrane-depleted nuclei were subjected to short term (20 min) exposure to µ-calpain, similar gel shift together with an increase in PI3K-C2beta activity was observed, when compared with the nuclei harvested 20 h after partial hepatectomy. Moreover, the above-mentioned gel shift and increase in PI3K-C2beta activity could be prevented by the calpain inhibitor calpeptin. The data presented in this report show that, in the membrane-depleted nuclei during the compensatory liver growth, there is an increase in PtdIns(3)P formation as a result of PI3K-C2beta activation, which may be a calpain-mediated event.

* This work was supported by the Ministry of Science of the Republic of Croatia (to H. B.), by a Fogarty International Research Collaboration Award (to A. P. F. and H. B.), and by the AIRC (to S. V.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

** To whom correspondence should be addressed: Zavod za Fiziologiju, Medicinski Fakultet, Sveuciliste u Zagrebu, Salata 3, P. O. Box 978, Zagreb 10,001, Croatia. Tel.: 385-1-4590-260; Fax: 385-1-4590-207; E-mail: hrvoje.banfic@zg.tel.hr.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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