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Molecular and Cellular Biology, November 2000, p. 8047-8058, Vol. 20, No. 21
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of Novel Saccharomyces cerevisiae Proteins with Nuclear Export Activity: Cell Cycle-Regulated Transcription Factor Ace2p Shows Cell Cycle-Independent Nucleocytoplasmic Shuttling

Torben Heick Jensen,1 Megan Neville,1,dagger Jean Christophe Rain,2,Dagger Terri McCarthy,1 Pierre Legrain,2,Dagger and Michael Rosbash1,*

Howard Hughes Medical Institute, Department of Biology, Brandeis University, Waltham, Massachusetts 02454,1 and Unité de Génétique des Interactions Macromoléculaires- CNRS URA1300, Institut Pasteur, 75524 Paris Cedex 15, France2

Received 6 April 2000/Returned for modification 9 May 2000/Accepted 1 August 2000

Nuclear export of proteins containing leucine-rich nuclear export signals (NESs) is mediated by the NES receptor CRM1/Crm1p. We have carried out a yeast two-hybrid screen with Crm1p as a bait. The Crm1p-interacting clones were subscreened for nuclear export activity in a visual assay utilizing the Crm1p-inhibitor leptomycin B (LMB). This approach identified three Saccharomyces cerevisiae proteins not previously known to have nuclear export activity. These proteins are the 5' RNA triphosphatase Ctl1p, the cell cycle-regulated transcription factor Ace2p, and a protein encoded by the previously uncharacterized open reading frame YDR499W. Mutagenesis analysis show that YDR499Wp contains an NES that conforms to the consensus sequence for leucine-rich NESs. Mutagenesis of Ctl1p and Ace2p were unable to identify specific NES residues. However, a 29-amino-acid region of Ace2p, rich in hydrophobic residues, contains nuclear export activity. Ace2p accumulates in the nucleus at the end of mitosis and activates early-G1-specific genes. We now provide evidence that Ace2p is nuclear not only in late M-early G1 but also during other stages of the cell cycle. This feature of Ace2p localization explains its ability to activate genes such as CUP1, which are not expressed in a cell cycle-dependent manner.

* Corresponding author. Mailing address: Department of Biology, Brandeis University, 415 South St., Waltham, MA 02454. Phone: (781) 736-3160. Fax: (781) 736-3164. E-mail: rosbash{at}brandeis.edu.

dagger Present address: University of Glasgow, CRC Beatson Laboratories, Garscube Estate, Glasgow G61 1BD, United Kingdom.

Dagger Present address: Hybrigenics, Paris 75012, France.

Molecular and Cellular Biology, November 2000, p. 8047-8058, Vol. 20, No. 21
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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