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

We determined the times when the nuclear membrane, nuclear pore complex (NP C) 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 chr omosomes revealed that nuclear membranes reassembled around chromosomes by 5 minutes after the onset of anaphase (early telophase) whereas nuclear imp ort function was recovered later, at 8 minutes. GFP-tagged emerin also accu mulated 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 ti mes 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 recov ery of nuclear import activity.