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PMID: 7822421 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Depletion of calcium from the lumen of endoplasmic reticulum reversibly inhibits passive diffusion and signal-mediated transport into the nucleus.

The Journal of cell biology ·Vol. 128 ·No. 1-2 ·1995-01-00 ·Pages 5-14

Greber UF, Gerace L

Abstract

Nuclear pore complexes provide channels for molecular transport across the nuclear envelope. Translocation of most proteins and RNAs through the pore complex is mediated by signal- and ATP-dependent mechanisms, while transport of small molecules is accomplished by passive diffusion. We report here that depletion of calcium from the lumen of the endoplasmic reticulum and nuclear envelope with ionophores or the calcium pump inhibitor thapsigargin rapidly and potently inhibits signal mediated transport of proteins into the nucleus. Lumenal calcium depletion also inhibits passive diffusion through the pore complex. Signal-mediated protein import and passive diffusion are rapidly restored when the drugs depleting lumenal calcium are removed and cells are incubated at 37 degrees C in calcium-containing medium. These results indicate that loss of calcium from the lumen of the endoplasmic reticulum and nuclear envelope reversibly affects properties of pore complex components located on the nuclear/cytoplasmic membrane surfaces, and they provide direct functional evidence for conformational flexibility of the pore complex. These methods will be useful for achieving reversible inhibition of nucleocytoplasmic trafficking for in vivo functional studies, and for studying the structure of the passive diffusion channel(s) of the pore complex.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Calcimycin/administration & dosage,pharmacology Calcium/physiology Calcium-Transporting ATPases/antagonists & inhibitors Cell Line Cell Nucleus/metabolism Diffusion Egtazic Acid/pharmacology Endoplasmic Reticulum/metabolism HeLa Cells Humans Kidney Microinjections Nuclear Envelope/metabolism Nuclear Proteins/metabolism Rats Signal Transduction/drug effects Terpenes/administration & dosage,pharmacology Thapsigargin
Chemicals
Nuclear Proteins Terpenes Calcimycin Egtazic Acid Thapsigargin Adenosine Triphosphate Calcium-Transporting ATPases Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Greber U F
Yale University School of Medicine, Department of Cell Biology, New Haven, Connecticut 06520.
Gerace L
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.128.1.5
Published
1995-01-00
Pages
5-14
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120336
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.128.1.5 PMC Full Text View journal details
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