4.4 Article

Visualization of actin filaments and monomers in somatic cell nuclei

Journal

MOLECULAR BIOLOGY OF THE CELL
Volume 24, Issue 7, Pages 982-994

Publisher

AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E12-09-0685

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Funding

  1. National Institutes of Health [5R01GM061010-12, R01 CA096840]
  2. National Science Foundation Predoctoral Fellowship
  3. Genentech Fellowship

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In addition to its long-studied presence in the cytoplasm, actin is also found in the nuclei of eukaryotic cells. The function and form (monomer, filament, or noncanonical oligomer) of nuclear actin are hotly debated, and its localization and dynamics are largely unknown. To determine the distribution of nuclear actin in live somatic cells and evaluate its potential functions, we constructed and validated fluorescent nuclear actin probes. Monomeric actin probes concentrate in nuclear speckles, suggesting an interaction of monomers with RNA-processing factors. Filamentous actin probes recognize discrete structures with submicron lengths that are excluded from chromatin-rich regions. In time-lapse movies, these actin filament structures exhibit one of two types of mobility: 1) diffusive, with an average diffusion coefficient of 0.06-0.08 mu m(2)/s, or (2) subdiffusive, with a mobility coefficient of 0.015 mu m(2)/s. Individual filament trajectories exhibit features of particles moving within a viscoelastic mesh. The small size of nuclear actin filaments is inconsistent with a role in micron-scale intranuclear transport, and their localization suggests that they do not participate directly in chromatin-based processes. Our results instead suggest that actin filaments form part of a large, viscoelastic structure in the nucleoplasm and may act as scaffolds that help organize nuclear contents.

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