Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation

D D Newmeyer, J M Lucocq, T R Bürglin, E M De Robertis

Research output: Contribution to journalArticlepeer-review

133 Citations (Scopus)

Abstract

DNA (from bacteriophage lambda or Xenopus) is assembled into nucleus-like structures when mixed with an extract from Xenopus eggs. Electron microscopy shows that these in vitro-reconstituted nuclei possess complete double membranes; some, but not all, nuclei have pore complexes. Extracts depleted of their endogenous ATP (by addition of ATPases) cannot assemble nuclear envelopes visible by phase-contrast microscopy. Once synthetic nuclei are assembled, however, they are stable when ATP is subsequently depleted, although their chromatin becomes condensed. About one-fourth of the nuclei assembled in vitro from lambda DNA accumulate nuclear proteins such as nucleoplasmin. ATP depletion blocks nucleoplasmin accumulation both in vitro, in pre-assembled synthetic nuclei, and in vivo, in the nucleus of microinjected oocytes. However, nucleoplasmin previously accumulated by reconstituted nuclei or by the germinal vesicle in microinjected oocytes is retained after ATP depletion.
Original languageEnglish
Pages (from-to)501-10
Number of pages10
JournalEMBO Journal
Volume5
Issue number3
Publication statusPublished - Mar 1986

Keywords

  • Adenosine Triphosphate
  • Animals
  • Autoradiography
  • Biological Transport
  • Cell Nucleus
  • DNA
  • Female
  • Iodine Radioisotopes
  • Kinetics
  • Microscopy, Electron
  • Nuclear Envelope
  • Nuclear Proteins
  • Nucleoplasmins
  • Nucleoproteins
  • Ovum
  • Phosphoproteins
  • Xenopus

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