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PMID: 9724751 Published · ppublish English Journal Article

Reconstitution of HIV-1 rev nuclear export: independent requirements for nuclear import and export.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 95 ·No. 18 ·1998-09-01 ·Pages 10608-13

Love DC, Sweitzer TD, Hanover JA

Abstract

The Rev protein of HIV-1 actively shuttles between nucleus and cytoplasm and mediates the export of unspliced retroviral RNAs. The localization of shuttling proteins such as Rev is controlled by the relative rates of nuclear import and export. To study nuclear export in isolation, we generated cell lines expressing a green fluorescent protein-labeled chimeric protein consisting of HIV-1 Rev and a hormone-inducible nuclear localization sequence. Steroid removal switches off import thus allowing direct visualization of the Rev export pathway in living cells. After digitonin permeabilization of these cells, we found that a functional nuclear export sequence (NES), ATP, and fractionated cytosol were sufficient for nuclear export in vitro. Nuclear pore-specific lectins and leptomycin B were potent export inhibitors. Nuclear export was not inhibited by antagonists of calcium metabolism that block nuclear import. These data further suggest that nuclear pores do not functionally close when luminal calcium stores are depleted. The distinct requirements for nuclear import and export argue that these competing processes may be regulated independently. This system should have wide applicability for the analysis of nuclear import and export.

MeSH Terms
Adenosine Triphosphate/metabolism Biological Transport Calcium/metabolism Cell Nucleus/metabolism Gene Products, rev/metabolism HIV-1/metabolism HeLa Cells Humans Hydrolysis rev Gene Products, Human Immunodeficiency Virus
Chemicals
Gene Products, rev rev Gene Products, Human Immunodeficiency Virus Adenosine Triphosphate Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Love D C
Laboratory of Cell Biochemistry and Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0851, USA.
Sweitzer T D
Hanover J A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.95.18.10608
Published
1998-09-01
Pages
10608-13
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC27942
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.95.18.10608 PMC Full Text View journal details
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