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The Nucleus pp 207-223 | Cite as

Nuclear Envelope Formation In Vitro: A Sea Urchin Egg Cell-Free System

  • Richard D. Byrne
  • Vanessa Zhendre
  • Banafshé Larijani
  • Dominic L. Poccia
Part of the Methods in Molecular Biology book series (MIMB, volume 464)

Abstract

The formation of the nuclear envelope (NE) typically occurs once during every mitotic cycle in somatic cells, and also around the sperm nucleus following fertilization. Much of our understanding of NE assembly has been derived from systems modeling the latter event in vitro. In these systems, demembranated sperm nuclei are combined with fertilized egg cytoplasmic extracts and an ATP-regenerating system and in a multistep process they form the functional double bilayer of the NE. Using a system that we developed from sea urchin gametes, we have demonstrated that NE assembly is regulated by membrane vesicles in a spatial and temporal fashion, emphasizing the roles of phosphoinositides, particularly phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), diacylglycerols (DAG), and lipid-modifying enzymes in NE assembly.

Keywords

Nuclear envelope Sea urchin Phosphatidylinositol Phosphatidylcholine Cholesterol Phospholipase C PtdIns(4;5)P2 Diacylglycerol Liquid NMR Liquid nuclear magnetic resonance 

Notes

Acknowledgments

This work was supported by an Amherst College Faculty Research Award of the H. Axel Schupf ‘57 Fund for Intellectual Life (DLP) and core funding from Cancer Research UK (BL).

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Copyright information

© Humana Press, a part of Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • Richard D. Byrne
    • 1
  • Vanessa Zhendre
    • 1
  • Banafshé Larijani
    • 1
  • Dominic L. Poccia
    • 2
  1. 1.Cell Biophysics Laboratory, Cancer Research UK (CR-UK)London Research Institute (LRI)LondonUK
  2. 2.Department of BiologyAmherst CollegeAmherstUSA

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