Aldosterone activates the nuclear pore transporter in cultured kidney cells imaged with atomic force microscopy.
Nuclear pore complexes (NPC), located in the nuclear envelope, functionally connect the cell nucleus with the cytoplasm and serve as a crucial pathway for macromolecule exchange. A Madin-Darby canine kidney (MDCK) clone that resembles principal cells of the collecting duct was shown recently to respond to sustained aldosterone exposure with a significant increase in the NPC number per nucleus. The present study elucidates the molecular nature of the NPC pathway and its regulation by aldosterone applying atomic force microscopy. We imaged individual NPC in situ and searched for a putative so-called transporter in the NPC centre. In aldosterone-depleted cells we found numerous macromolecules docked to individual NPC waiting for translocation into the nucleoplasm (standby mode=inactive pore). In contrast, in aldosterone-treated cells NPC were frequently found free of macromolecules, indicating that the translocation process kept pace with docking under hormone-stimulated conditions (transport mode=active pore). In the NPC centre we detected a ring-like structure with a central invagination. We assume that the ring is the putative transporter and that the invagination is the channel entrance used for translocation of macromolecules. Transporters were found in open and closed configurations. In conclusion, the results provide evidence for the existence of a nuclear transporter as part of the translocation machinery of an individual NPC. Aldosterone increases the activity of the nuclear transporter and thus facilitates steroid-mediated gene expression.
Folprecht
G
G
Department of Physiology, University of Würzburg, Röntgenring 9, D-97070 Würzburg, Germany.
Schneider
S
S
Oberleithner
H
H
D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D001693:Biological Transport, Active; D002478:Cells, Cultured; D004285:Dogs; D007091:Image Processing, Computer-Assisted; D007685:Kidney Tubules, Collecting / Q000187:drug effects* / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D009685:Nuclear Envelope / Q000187:drug effects* / Q000648:ultrastructure
D000450:Aldosterone