Current research in our group focuses on the vertebrate nuclear pore complex (NPC). The main biological questions we are investigating are: (1) How is this supramolecular machine assembled and integrated into the double membranes surrounding the nucleus; (2) The mechanism for targeting and translocation through the NPC of large cellular cargoes and viruses. To study these topics we rely on the cell-free nuclear reconstitution system, derived from frog egg extracts and on genetic manipulation of mammalian cells.
The main source of biological material for our experiments are unfertilized frog eggs, shown on the right. These eggs are used to produce fractionated cell-free extracts, which allow us to assemble functional nuclei in the test-tube. The nuclei can be directly visualized by fluorescent, confocal or electron microscopy.
Direct immunofluorescent staining of NPCs on the surface of a reconstituted nucleus. Staining by an affinity-purified antibody directed against the pore-membrane protein POM121. See more about the functional roles of this protein in Shaulov et al., J. Cell Science, 2011.
A major research goal in this field of cell biology is to deduce a step-by-step molecular mechanism for the entire process of NPC biogenesis and to place NPC and nuclear envelope assembly in the wider context of cell-cycle progression. For a global view of the mechanistic questions driving this field of research, see: Wandke and Kutay, Cell 152, 1222-5, 2013.
Other projects in the lab involve the targeting of intact proteasome particles to the nuclear compartment, the impact of heat-shock and stress on nuclear transport, the viral-host interactions of Herpes Simplex Virus 1 (HSV1) and a rare genetic disease affecting a nuclear pore component in humans.