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Molecular and Cellular Biology, July 2000, p. 4562-4571, Vol. 20, No. 13
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

RanGTP-Binding Protein NXT1 Facilitates Nuclear Export of Different Classes of RNA In Vitro

Batool Ossareh-Nazari,1 Christèle Maison,1,dagger Ben E. Black,2 Lyne Lévesque,2 Bryce M. Paschal,2 and Catherine Dargemont1,*

Laboratoire de Transport Nucléocytoplasmique, Unité Mixte de Recherche 144, Institut Curie-CNRS, 75248 Paris Cedex 05, France,1 and Center for Cell Signaling, Department of Biochemistry and Molecular Genetics, University of Virginia Health Sciences Center, Charlottesville, Virginia 229082

Received 14 December 1999/Returned for modification 20 January 2000/Accepted 11 April 2000

To better characterize the mechanisms responsible for RNA export from the nucleus, we developed an in vitro assay based on the use of permeabilized HeLa cells. This new assay supports nuclear export of U1 snRNA, tRNA, and mRNA in an energy- and Xenopus extract-dependent manner. U1 snRNA export requires a 5' monomethylated cap structure, the nuclear export signal receptor CRM1, and the small GTPase Ran. In contrast, mRNA export does not require the participation of CRM1. We show here that NXT1, an NTF2-related protein that binds directly to RanGTP, strongly stimulates export of U1 snRNA, tRNA, and mRNA. The ability of NXT1 to promote export is dependent on its capacity to bind RanGTP. These results support the emerging view that NXT1 is a general export factor, functioning on both CRM1-dependent and CRM1-independent pathways of RNA export.

* Corresponding author. Mailing address: Laboratoire de Transport Nucléocytoplasmique, Unité Mixte de Recherche 144 Institut Curie-CNRS, 26 rue d'Ulm, 75248 Paris Cedex 05, France. Phone: 0033 1 42346366. Fax: 0033 1 42346367. E-mail: dargemon{at}curie.fr.

dagger Present address: Laboratoire de la Dynamique Nucléaire et de la Plasticité du Genome, Unité Mixte de Recherche 218 Institut Curie-CNRS, 75248 Paris Cedex 05, France.

Molecular and Cellular Biology, July 2000, p. 4562-4571, Vol. 20, No. 13
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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