Diffusion of large molecules into assembling nuclei revealed using an optical highlighting technique.
The nuclear envelope (NE) defines the nuclear compartment, and nuclear pore complexes (NPCs) on the NE form aqueous passages through which small water-soluble molecules can passively diffuse. It is well known that proteins smaller than 50 kDa can diffuse though NPCs, whereas proteins larger than 60 kDa rarely enter by passive diffusion. Little, however, is known about how this size cutoff develops as the NE reassembles and the nucleus expands. In 1987, a well-known study identified an efficient mechanism by which large diffusing proteins (> 60 kDa) were excluded from the reassembling nucleus after mitosis. Since then, it has been generally accepted that after mitosis, newly formed nuclei completely exclude all proteins except those that are initially bound to the mitotic chromosomes and those that are selectively imported through NPCs. Here, the tetrameric complex of the photoconvertible fluorescent protein KikGR ( approximately 103 kDa) was optically highlighted in the cytoplasm and followed to examine its entry into nuclei. Remarkably, highlighted complexes efficiently entered newly assembled nuclei during an approximately 20-min period after the completion of cytokinesis. Because KikGR contains no known nuclear-localization or chromosome-binding sequences, our results indicate the diffusion barrier is less restrictive during nuclear reassembly.
Shimozono
Satoshi
S
Laboratory for Cell Function Dynamics, Advanced Technology Development Group, Brain Science Institute, Institute of Physical and Chemical Research, Saitama, Japan.
Tsutsui
Hidekazu
H
Miyawaki
Atsushi
A
D021581:Active Transport, Cell Nucleus; D000818:Animals; D037421:Anthozoa; D002455:Cell Division / Q000502:physiology*; D002463:Cell Membrane Permeability; D002467:Cell Nucleus / Q000378:metabolism*; D048749:Cytokinesis / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism; D004058:Diffusion; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008938:Mitosis; D008958:Models, Molecular; D009154:Mutation; D055095:Optics and Photonics / Q000379:methods*; D013997:Time Factors
D049452:Green Fluorescent Proteins