Nuclear pore ion channel activity in live syncytial nuclei.
Nuclear pore complexes (NPCs) are important nanochannels for the control of gene activity and expression. Most of our knowledge of NPC function has been derived from isolated nuclei and permeabilized cells in cell lysates/extracts. Since recent patch-clamp work has challenged the dogma that NPCs are freely permeable to small particles, a preparation of isolated living nuclei in their native liquid environment was sought and found: the syncytial nuclei in the water of the coconut Cocos nucifera. These nuclei have all properties of NPC-mediated macromolecular transport (MMT) and express foreign green fluorescent protein (GFP) plasmids. They display chromatin movement, are created by particle aggregation or by division, can grow by throwing filaments to catch material, etc. This study shows, for the first time, that living NPCs engaged in MMT do not transport physiological ions - a phenomenon that explains observations of nucleocytoplasmic ion gradients. Since coconuts are inexpensive (less than US$1/nut per litre), this robust preparation may contribute to our understanding of NPCs and cell nucleus and to the development of biotechnologies for the production of DNA, RNA and proteins.
Bustamante
Jose Omar
JO
The Nuclear Physiology Lab-ITP-UNIT, The Millenium Institute of Nanosciences, Brazilian Ministry of Science and Technology, Praia Aruana, Aracaju, Sergipe 49037-610, Brazil. jobustamante@yahoo.com
D002467:Cell Nucleus / Q000502:physiology* / Q000648:ultrastructure; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure; D003059:Cocos / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004594:Electrophysiology; D049452:Green Fluorescent Proteins; D007473:Ion Channels / Q000502:physiology* / Q000648:ultrastructure; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D018408:Patch-Clamp Techniques; D010539:Permeability; D010957:Plasmids; D012313:RNA / Q000096:biosynthesis
D002843:Chromatin; D007473:Ion Channels; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D012313:RNA; D004247:DNA