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PMID: 11149926 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Calreticulin Is a receptor for nuclear export.

The Journal of cell biology ·Vol. 152 ·No. 1 ·2001-01-08 ·Pages 127-40

Holaska JM, Black BE, Love DC, Hanover JA, Leszyk J, Paschal BM

Abstract

In previous work, we used a permeabilized cell assay that reconstitutes nuclear export of protein kinase inhibitor (PKI) to show that cytosol contains an export activity that is distinct from Crm1 (Holaska, J.M., and B.M. Paschal. 1995. Proc. Natl. Acad. Sci. USA. 95: 14739-14744). Here, we describe the purification and characterization of the activity as calreticulin (CRT), a protein previously ascribed to functions in the lumen of the ER. We show that cells contain both ER and cytosolic pools of CRT. The mechanism of CRT-dependent export of PKI requires a functional nuclear export signal (NES) in PKI and involves formation of an export complex that contains RanGTP. Previous studies linking CRT to downregulation of steroid hormone receptor function led us to examine its potential role in nuclear export of the glucocorticoid receptor (GR). We found that CRT mediates nuclear export of GR in permeabilized cell, microinjection, and transfection assays. GR export is insensitive to the Crm1 inhibitor leptomycin B in vivo, and it does not rely on a leucine-rich NES. Rather, GR export is facilitated by its DNA-binding domain, which is shown to function as an NES when transplanted to a green fluorescent protein reporter. CRT defines a new export pathway that may regulate the transcriptional activity of steroid hormone receptors.

MeSH Terms
Active Transport, Cell Nucleus Calcium-Binding Proteins/metabolism Calreticulin Carrier Proteins/metabolism Cell Nucleus/metabolism Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors Cytosol/metabolism Fatty Acids, Unsaturated/pharmacology HeLa Cells Humans Intracellular Signaling Peptides and Proteins Receptors, Cytoplasmic and Nuclear/metabolism Receptors, Glucocorticoid/metabolism Ribonucleoproteins/metabolism ran GTP-Binding Protein/metabolism
Chemicals
Calcium-Binding Proteins Calreticulin Carrier Proteins Fatty Acids, Unsaturated Intracellular Signaling Peptides and Proteins Receptors, Cytoplasmic and Nuclear Receptors, Glucocorticoid Ribonucleoproteins protein kinase modulator Cyclic AMP-Dependent Protein Kinases ran GTP-Binding Protein leptomycin B
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Holaska J M
Center for Cell Signaling, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA.
Black B E
Love D C
Hanover J A
Leszyk J
Paschal B M
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.152.1.127
Published
2001-01-08
Pages
127-40
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2193655
Subset
IM
Grants
NIGMS NIH HHS · GM58639-01 · United States
External Links
PubMed source DOI: 10.1083/jcb.152.1.127 PMC Full Text View journal details
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