jasplakinolide and 4-4-difluoro-4-bora-3a-4a-diaza-s-indacene

There exists abundant molecular and ultra-structural evidence to suggest that cytoplasmic actin can physically interact with the nuclear envelope (NE) membrane system. However, this interaction has yet to be characterised in living interphase cells.. Using a fluorescent conjugate of the actin binding drug cytochalasin D (CD-BODIPY) we provide evidence that polymerising actin accumulates in vicinity to the NE. In addition, both transiently expressed fluorescent actin and cytoplasmic micro-injection of fluorescent actin resulted in accumulation of actin at the NE-membrane. Consistent with the idea that the cytoplasmic phase of NE-membranes can support this novel pool of perinuclear actin polymerisation we show that isolated, intact, differentiated primary hepatocyte nuclei support actin polymerisation in vitro. Further this phenomenon was inhibited by treatments hindering steric access to outer-nuclear-membrane proteins (e.g. wheat germ agglutinin, anti-nesprin and anti-nucleoporin antibodies).. We conclude that actin polymerisation occurs around interphase nuclei of living cells at the cytoplasmic phase of NE-membranes.

Topics: Actins; Animals; Binding Sites; Biopolymers; Boron Compounds; Bridged Bicyclo Compounds, Heterocyclic; Circular Dichroism; Cytochalasin D; Depsipeptides; Fluorescent Dyes; HeLa Cells; Humans; Liver; Nuclear Envelope; Rabbits; Rats; Thiazoles; Thiazolidines