title: Formation of membrane networks in vitro by kinesin-driven microtubule movement.
abstract: Certain intracellular organelles such as the endoplasmic reticulum (Terasaki, M., L. B. Chen, and K. Fujiwara. 1986. J. Cell Biol. 103:1557-1568) and lysosomes (Swanson, J., A. Bushnell, and S. C. Silverstein. Proc. Natl. Acad. Sci. USA. 84:1921-1925) form tubular networks that are closely aligned with microtubules. Here we describe the formation of polygonal networks composed of interconnected membrane tubules that occurs when a preparation of microtubule affinity-purified squid kinesin is combined with microtubules and ATP on a glass surface. The membrane, which is a minor contaminant in the microtubule affinity-purified kinesin preparation, binds to microtubules translocating along kinesin-coated glass surfaces. Force exerted by kinesin upon the microtubule is transmitted to the membrane and a tubular extension of the membrane is produced. As the membrane tubule elongates, membrane tension exerts an opposing force upon the translocating microtubule that can alter its direction of movement by dissociating or partially dissociating the microtubule from the kinesin-coated surface. Membrane tubules that come in contact appear to fuse with one another, and thus give rise to two-dimensional polygonal networks of tubules that have similar features to endoplasmic reticulum networks in cells. Artificial liposomes composed of dimyristoylphosphatidylcholine and yolk phosphatidylglycerol also form stable tubular structures when subjected to shear forces, but do not interact with microtubules or form polygonal networks, suggesting that such phenomena may require membrane-associated proteins. These findings indicate that kinesin generates sufficient force to form tubular membrane extensions in vitro and suggest that this microtubule-based motility protein may also be responsible for creating tubular membrane networks within cells.
mesh_terms: D000818:Animals; D002474:Cell-Free System; D049832:Decapodiformes; D004721:Endoplasmic Reticulum / Q000502:physiology / Q000648:ultrastructure; D007425:Intracellular Membranes / Q000502:physiology* / Q000648:ultrastructure; D016547:Kinesins; D008081:Liposomes; D008870:Microtubules / Q000502:physiology*; D009024:Morphogenesis; D009068:Movement; D009419:Nerve Tissue Proteins / Q000502:physiology*
chemical_list: D008081:Liposomes; D009419:Nerve Tissue Proteins; D016547:Kinesins