Patch clamp and atomic force microscopy demonstrate TATA-binding protein (TBP) interactions with the nuclear pore complex

1995 ◽  
Vol 146 (3) ◽  
Author(s):  
J.O. Bustamante ◽  
A. Liepins ◽  
R.A. Prendergast ◽  
J.A. Hanover ◽  
H. Oberleithner
Keyword(s):  
Atomic Force Microscopy ◽  
Patch Clamp ◽  
Nuclear Pore Complex ◽  
Binding Protein ◽  
Tata Binding Protein ◽  
Nuclear Pore ◽  
Force Microscopy ◽  
Atomic Force

Immunolocalization of lamins and nuclear pore complex proteins by atomic force microscopy

1995 ◽  
Vol 430 (5) ◽  
pp. 795-801 ◽  
Author(s):  
Stefan Schneider ◽  
Gunnar Folprecht ◽  
Georg Krohne ◽  
Hans Oberleithner
Keyword(s):  
Atomic Force Microscopy ◽  
Nuclear Pore Complex ◽  
Nuclear Pore ◽  
Force Microscopy ◽  
Atomic Force

Towards monitoring transport of single cargos across individual nuclear pore complexes by time-lapse atomic force microscopy

2010 ◽  
Vol 171 (2) ◽  
pp. 154-162 ◽  
Author(s):  
Ning-Ping Huang ◽  
Mike Stubenrauch ◽  
Joachim Köser ◽  
Nicole Taschner ◽  
Ueli Aebi ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Time Lapse ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Force Microscopy ◽  
Atomic Force ◽  
Pore Complexes

Route of steroid-activated macromolecules through nuclear pores imaged with atomic force microscopy

2003 ◽  
Vol 31 (1) ◽  
pp. 71-75 ◽  
Author(s):  
H. Oberleithner ◽  
C. Schäfer ◽  
V. Shahin ◽  
L. Albermann
Keyword(s):  
Atomic Force Microscopy ◽  
Molecular Mass ◽  
Intact Cell ◽  
Rnase A ◽  
Nuclear Pore ◽  
Xenopus Laevis Oocyte ◽  
Cell Nuclei ◽  
Homogeneous Population ◽  
Force Microscopy ◽  
Atomic Force

In eukaryotic cells, two concentric membranes, the nuclear envelope (NE), separate the nucleus from the cytoplasm. The NE is punctured by nuclear pore complexes (NPCs; molecular mass 120 MDa) that serve as regulated pathways for macromolecules entering and leaving the nuclear compartment. Transport across NPCs occurs through central channels. Such import and export of macromolecules through individual NPCs can be elicited in the Xenopus laevis oocyte by injecting the mineralocorticoid aldosterone and can be visualized with atomic force microscopy. The electrical NE resistance in intact cell nuclei can be measured in parallel. Resistance increases when macromolecules are engaged with the NPC. This article describe six observations made from these experiments and the conclusions that can be drawn from them. (i) A homogeneous population of macromolecules (approx. 100 kDa) attaches to the cytoplasmic face of the NPC 2 min after aldosterone injection. They are most likely to be aldosterone receptors. After a few minutes, they have disappeared. (ii) Large plugs (approx. molecular mass 1 MDa) appear in the central channels 20 min after hormone injection. They are most likely to be ribonucleoproteins exiting the nucleus. (iii) Electrical resistance measurements in isolated nuclei reveal transient electrical NE resistance peaks: an early (2 min) peak and a late (20 min) peak. Electrical peaks reflect macromolecule interaction with the NPC. (iv) Spironolactone blocks both the early and late peaks. This indicates that classic aldosterone receptors are involved in the pregenomic (early) and post-genomic (late) responses. (v) Actinomycin D and, independently, RNase A block the late electrical peak, confirming that plugs are genomic in nature. (vi) Intracellular calcium chelation blocks both early and late electrical peaks. Thus, the release of calcium from internal stores, which is known to be the first intracellular signal in response to aldosterone, is a prerequisite for the late genomic response.

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