Actin-filled nuclear invaginations indicate degree of cell de-differentiation.
For years the existence of nuclear actin has been heavily debated, but recent data have clearly demonstrated that actin, as well as actin-binding proteins (ABPs), are located in the nucleus. We examined live EGFP-actin-expressing cells using confocal microscopy and saw the presence of structures strongly resembling actin filaments in the nuclei of MDA-MB-231 human mammary epithelial tumor cells. Many nuclei had more than one of these filamentous structures, some of which appeared to cross the entire nucleus. Extensive analysis, including fluorescence recovery after photobleaching (FRAP), showed that all EGFP-actin in the nucleus is monomeric (G-actin) rather than filamentous (F-actin) and that the apparent filaments seen in the nucleus are invaginations of cytoplasmic monomeric actin. Immunolocalization of nuclear pore complex proteins shows that similar invaginations are seen in cells that are not overexpressing EGFP-actin. To determine whether there is a correlation between increased levels of invagination in the cell nuclei and the state of de-differentiation of the cell, we examined a variety of cell types, including live Xenopus embryonic cells. Cells that were highly de-differentiated, or cancerous, had an increased incidence of invagination, while cells that were differentiated had few nuclear invaginations. The nuclei of embryonic cells that were not yet differentiated underwent multiple shape changes throughout interphase, and demonstrated numerous transient invaginations of varying sizes and shapes. Although the function of these actin-filled invaginations remains speculative, their presence correlates with cells that have increased levels of nuclear activity.
Johnson
Nicole
N
Advanced Light Source Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Krebs
Matthew
M
Boudreau
Rosanne
R
Giorgi
Gisele
G
LeGros
Mark
M
Larabell
Carolyn
C
D008841:Actin Cytoskeleton / Q000648:ultrastructure; D000199:Actins / Q000032:analysis*; D000818:Animals; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry / Q000648:ultrastructure*; D003593:Cytoplasm / Q000737:chemistry / Q000648:ultrastructure; D005260:Female; D036681:Fluorescence Recovery After Photobleaching; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D009685:Nuclear Envelope / Q000737:chemistry / Q000648:ultrastructure
D000199:Actins; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins