Live fluorescence imaging reveals early recruitment of emerin, LBR, RanBP2, and Nup153 to reforming functional nuclear envelopes

Tokuko Haraguchi, Takako Koujin, Tomohiro Hayakawa, Toru Kaneda, Chihiro Tsutsumi, Naoko Imamoto, Chihiro Akazawa, Jun Sukegawa, Yoshihiro Yoneda, Yasushi Hiraoka

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150 Citations (Scopus)

Abstract

We determined the times when the nuclear membrane, nuclear pore complex (NPC) components, and nuclear import function were recovered during telophase in living HeLa cells. Simultaneous observation of fluorescently-labeled NLS-bearing proteins, lamin B receptor (LBR)-GFP, and Hoechst33342-stained chromosomes revealed that nuclear membranes reassembled around chromosomes by 5 minutes after the onset of anaphase (early telophase) whereas nuclear import function was recovered later, at 8 minutes. GFP-tagged emerin also accumulated on chromosomes 5 minutes after the onset of anaphase. Interestingly, emerin and LBR initially accumulated at distinct, separate locations, but then became uniform 8 minutes after the onset of anaphase, concurrent with the recovery of nuclear import function. We further determined the timing of NPC assembly by immunofluorescence staining of cells fixed at precise times after the onset of anaphase. Taken together, these results showed that emerin, LBR, and several NPC components (RanBP2, Nup153, p62), but not Tpr, reconstitute around chromosomes very early in telophase prior to the recovery of nuclear import activity.

Original languageEnglish
Pages (from-to)779-794
Number of pages16
JournalJournal of cell science
Volume113
Issue number5
Publication statusPublished - 2000

Keywords

  • Chromosome decondensation
  • Live analysis
  • Mitosis
  • Nuclear envelope
  • Nuclear transport

ASJC Scopus subject areas

  • Cell Biology

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    Haraguchi, T., Koujin, T., Hayakawa, T., Kaneda, T., Tsutsumi, C., Imamoto, N., Akazawa, C., Sukegawa, J., Yoneda, Y., & Hiraoka, Y. (2000). Live fluorescence imaging reveals early recruitment of emerin, LBR, RanBP2, and Nup153 to reforming functional nuclear envelopes. Journal of cell science, 113(5), 779-794.