Ran-mediated nuclear export of the von Hippel-Lindau tumor suppressor protein occurs independently of its assembly with cullin-2

Isabelle Groulx, Marie Eve Bonicalzi, Stephen Lee

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Inactivating mutations of the von Hippel-Lindau (VHL) tumor suppressor gene cause the VHL cancer syndrome and sporadic renal clear cell carcinoma. VHL engages in a nucleocytoplasmic shuttle, which is required for its function. Here, we pursue our investigation to identify mechanisms by which VHL-green fluorescent protein (VHL-GFP) is exported from the nucleus. We show that nuclear export of VHL-GFP in living cells requires ongoing RNA polymerase II activity, and is mediated by mechanisms that are temperature- sensitive and energy-dependent. In vitro nuclear export of VHL-GFP is inhibited by nuclear pore-specific lectins, requires ATP hydrolysis and polyadenylated mRNAs, and occurs with kinetics that are similar to those of proteins containing a nuclear export signal. Biochemical fractionation has revealed that nuclear export of VHL-GFP occurs by way of a Ran-dependent pathway. Size exclusion column chromatography and deletion mutant analysis suggest that VHL-GFP does not require assembly with one of its associated proteins, cullin-2, to engage in nuclear export. These results demonstrate that nuclear export of VHL-GFP is Ran-mediated and ATP hydrolysis-dependent. They also suggest that sequences outside the elongin C binding box may function as a nuclear export domain, potentially providing a novel role for this region of VHL frequently mutated in renal cell carcinoma.

Original languageEnglish (US)
Pages (from-to)8991-9000
Number of pages10
JournalJournal of Biological Chemistry
Volume275
Issue number12
DOIs
StatePublished - Mar 24 2000
Externally publishedYes

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Von Hippel-Lindau Tumor Suppressor Protein
Cullin Proteins
Cell Nucleus Active Transport
Cells
Hydrolysis
Adenosine Triphosphate
Column chromatography
Size exclusion chromatography
RNA Polymerase II
Fractionation
Green Fluorescent Proteins
Renal Cell Carcinoma
Lectins
Tumors
Genes
Nuclear Export Signals
von Hippel-Lindau Disease
Nuclear Pore
Messenger RNA
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ran-mediated nuclear export of the von Hippel-Lindau tumor suppressor protein occurs independently of its assembly with cullin-2. / Groulx, Isabelle; Bonicalzi, Marie Eve; Lee, Stephen.

In: Journal of Biological Chemistry, Vol. 275, No. 12, 24.03.2000, p. 8991-9000.

Research output: Contribution to journalArticle

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