Regulated Docking of Nuclear Membrane Vesicles to Vimentin Filaments during Mitosis

• Published in: The Journal of cell biology 1993-12, Vol.123 (6), p.1491-1505
• Main Authors: Maison, Christèle, Horstmann, Heinz, Georgatos, Spyros D.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Biological and medical sciences; Cell Compartmentation; Cell cycle, cell proliferation; Cell division; Cell Line; Cell membranes; Cell physiology; Cells; CHO cells; Cricetinae; Epithelial cells; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; HeLa cells; Immunohistochemistry; In Vitro Techniques; Intermediate Filaments - metabolism; Lamin Type B; Lamins; Membranes; Mitosis; Molecular and cellular biology; Nuclear Envelope - metabolism; Nuclear membrane; Nuclear Proteins - metabolism; Phosphorylation; Proteins; Vimentin; Vimentin - metabolism
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.123.6.1491

During mitosis, several types of intermediate-sized filaments (IFs) undergo an extensive remodelling in response to phosphorylation by cdc 2 and other protein kinases. However, unlike the nuclear lamins, the cytoplasmic IFs do not seem to follow a fixed disassembly stereotype and often retain their physical continuity without depolymerizing into soluble subunits. To investigate potential interactions between mitotically modified IFs and other cellular structures, we have examined prometaphase-arrested cells expressing the IF protein vimentin. We demonstrate here that vimentin filaments associate in situ and co-fractionate with a distinct population of mitotic vesicles. These vesicles carry on their surfaces nuclear lamin B, the inner nuclear membrane protein p58, and wheat germ agglutinin (WGA)-binding proteins. Consistent with a tight interaction between the IFs and the mitotic membranes, vimentin, nuclear lamin B, and a 180-kD WGA-binding protein are co-isolated when whole mitotic homogenates are incubated with antivimentin or anti-lamin B antibodies immobilized on magnetic beads. The vimentin-associated vesicles are essentially depleted of ER, Golgi and endosomal membrane proteins. The interaction of vimentin with lamin B-carrying membranes depends on phosphorylation and is weakened by dephosphorylation during nuclear reassembly in vitro. These observations reveal a novel interaction between IFs and cellular membranes and further suggest that the vimentin filaments may serve as a transient docking site for inner nuclear membrane vesicles during mitosis.