Studying nuclear disassembly in vitro using Xenopus egg extract

Meda M. Higa, Katharine S. Ullman, Amy J Prunuske

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Xenopus egg extract provides an extremely powerful approach in the study of cell cycle regulated aspects of nuclear form and function. Each egg contains enough membrane and protein components to support multiple rounds of cell division. Remarkably, incubation of egg extract with DNA in the presence of an energy regeneration system is sufficient to induce formation of a nuclear envelope around DNA. In addition, these in vitro nuclei contain functional nuclear pore complexes, which form de novo and are capable of supporting nucleocytoplasmic transport. Mitotic entry can be induced by the addition of recombinant cyclin to an interphase extract. This initiates signaling that leads to disassembly of the nuclei. Thus, this cell-free system can be used to decipher events involved in mitotic remodeling of the nuclear envelope such as changes in nuclear pore permeability, dispersal of membrane, and disassembly of the lamina. Both general mechanisms and individual players required for orchestrating these events can be identified via biochemical manipulation of the egg extract. Here, we describe a procedure for the assembly and disassembly of in vitro nuclei, including the production of Xenopus egg extract and sperm chromatin DNA.

Original languageEnglish (US)
Pages (from-to)284-290
Number of pages7
JournalMethods
Volume39
Issue number4
DOIs
StatePublished - Aug 1 2006

Fingerprint

Xenopus
Ovum
DNA
Nuclear Pore
Cells
Nuclear Envelope
Cyclins
Chromatin
Membrane Proteins
Cell Nucleus Active Transport
Cell-Free System
Membranes
Interphase
Cell Division
Spermatozoa
Regeneration
Permeability
Cell Cycle
In Vitro Techniques

Keywords

  • Disassembly
  • Lamina
  • Mitosis
  • Nuclear envelope breakdown
  • Nuclear membrane
  • Nuclear pore
  • Nucleus
  • Prophase
  • Sperm chromatin DNA
  • Xenopus egg extract

Cite this

Studying nuclear disassembly in vitro using Xenopus egg extract. / Higa, Meda M.; Ullman, Katharine S.; Prunuske, Amy J.

In: Methods, Vol. 39, No. 4, 01.08.2006, p. 284-290.

Research output: Contribution to journalArticle

Higa, MM, Ullman, KS & Prunuske, AJ 2006, 'Studying nuclear disassembly in vitro using Xenopus egg extract' Methods, vol. 39, no. 4, pp. 284-290. https://doi.org/10.1016/j.ymeth.2006.06.004
Higa, Meda M. ; Ullman, Katharine S. ; Prunuske, Amy J. / Studying nuclear disassembly in vitro using Xenopus egg extract. In: Methods. 2006 ; Vol. 39, No. 4. pp. 284-290.
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