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Originally published as MBC in Press, 10.1091/mbc.E05-07-0642 on August 24, 2005

Vol. 16, Issue 11, 5152-5162, November 2005

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Cdk1 and Okadaic Acid-sensitive Phosphatases Control Assembly of Nuclear Pore Complexes in Drosophila Embryos{boxv}

Evgeny A. Onischenko * {dagger}, Natalia V. Gubanova {ddagger}, Elena V. Kiseleva {ddagger}, and Einar Hallberg *

* Section of Life Sciences, Södertörns University College, SE-141 89 Huddinge, Sweden; {dagger} Department of Biosciences at Novum, Karolinska Institute, SE-141 86 Huddinge, Sweden; and {ddagger} Institute of Cytology and Genetics, 630090 Novosibirsk-90, Russia

Submitted July 18, 2005; Revised August 16, 2005; Accepted August 17, 2005
Monitoring Editor: Karsten Weis

Disassembly and reassembly of the nuclear pore complexes (NPCs) is one of the major events during open mitosis in higher eukaryotes. However, how this process is controlled by the mitotic machinery is not clear. To investigate this we developed a novel in vivo model system based on syncytial Drosophila embryos. We microinjected different mitotic effectors into the embryonic cytoplasm and monitored the dynamics of disassembly/reassembly of NPCs in live embryos using fluorescently labeled wheat germ agglutinin (WGA) or in fixed embryos using electron microscopy and immunostaining techniques. We found that in live embryos Cdk1 activity was necessary and sufficient to induce disassembly of NPCs as well as their cytoplasmic mimics: annulate lamellae pore complexes (ALPCs). Cdk1 activity was also required for keeping NPCs and ALPCs disassembled during mitosis. In agreement recombinant Cdk1/cyclin B was able to induce phosphorylation and dissociation of nucleoporins from the NPCs in vitro. Conversely, reassembly of NPCs and ALPCs was dependent on the activity of protein phosphatases, sensitive to okadaic acid (OA). Our findings suggest a model where mitotic disassembly/reassembly of the NPCs is regulated by a dynamic equilibrium of Cdk1 and OA-sensitive phosphatase activities and provide evidence that mitotic phosphorylation mediates disassembly of the NPC.

This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E05–07–0642) on August 24, 2005.

Abbreviations used: AL, annulate lamellae; ALPC, annulate lamellae pore complex; NE, nuclear envelope; NPC, nuclear pore complex; OA, okadaic acid; WGA, wheat germ agglutinin.

{boxv} The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Address correspondence to: Einar Hallberg (einar.hallberg{at}sh.se).




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