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Molecular and Cellular Biology, June 2004, p. 4824-4834, Vol. 24, No. 11
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.11.4824-4834.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Paired-Type Homeodomain Transcription Factors Are Imported into the Nucleus by Karyopherin 13

Jonathan E. Ploski,* Monee K. Shamsher,{dagger} and Aurelian Radu

The Carl C. Icahn Center for Gene Therapy and Molecular Medicine, The Mount Sinai School of Medicine, New York, New York 10029

Received 27 June 2003/ Returned for modification 23 September 2003/ Accepted 8 March 2004

We report that the paired homeodomain transcription factor Pax6 is imported into the nucleus by the Karyopherin ß family member Karyopherin 13 (Kap13). Pax6 was identified as a potential cargo for Kap13 by a yeast two-hybrid screen. Direct binding of Pax6 to Kap13 was subsequently confirmed by in vitro assays with recombinant proteins, and binding in vivo was shown by coimmunoprecipitation. Ran-dependent import of Pax6 by Kap13 was shown to occur by using a digitonin-permeabilized cells assay. Kap13 binds to Pax6 via a nuclear localization sequence (NLS), which is located within a segment of 80 amino acid residues that includes the homeodomain. Kap13 showed reduced binding to Pax6 when either region located at each end of the homeodomain (208 to 214 and 261 to 267) was deleted. The paired-type homeodomain transcription factor family includes more than 20 members. All members contain a region similar to the NLS found in Pax6 and are therefore likely to be imported by Kap13. We confirmed this hypothesis for Pax3 and Crx, which bind to and are imported by Kap13.

* Corresponding author. Mailing address: The Carl C. Icahn Center for Gene Therapy and Molecular Medicine, Box 1496, The Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029. Phone: (212) 241-3798. Fax: (212) 241-0738. E-mail: Jonathan.Ploski{at}mssm.edu.

{dagger} Present address: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.

Molecular and Cellular Biology, June 2004, p. 4824-4834, Vol. 24, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.11.4824-4834.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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