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MBC in Press, published online ahead of print May 11, 2005
Mol. Biol. Cell 10.1091/mbc.E04-11-0990

A more recent version of this article appeared on July 1, 2005
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Submitted on November 16, 2004
Revised on April 29, 2005
Accepted on May 3, 2005

Nuclear Import of TFIIB Is Mediated by Kap114p, a Karyopherin with Multiple Cargo Binding Domains

Jennifer L. Hodges,*{dagger} Jennifer H. Leslie,*{dagger} Nima Mosammaparast,* Yurong Guo,{ddagger}{sect} Jeffrey Shabanowitz,{ddagger} Donald F. Hunt,{ddagger} and Lucy F. Pemberton*

*Center for Cell Signaling, Department of Microbiology, University of Virginia Health Sciences Center; and {ddagger}Department of Chemistry, University of Virginia, Charlottesville, VA 22908

Monitoring Editor: Karsten Weis

Nuclear import and export is mediated by an evolutionarily conserved family of soluble transport factors, the karyopherins (referred to as importins and exportins). The yeast karyopherin Kap114p has previously been shown to import histones H2A and H2B, Nap1p, and a component of the preinitiation complex (PIC), TBP. Using a proteomic approach, we have identified several potentially new cargoes for Kap114p. These cargoes include another PIC component, the general transcription factor IIB or Sua7p, which interacted directly with Kap114p. Consistent with its role as a Sua7p import factor, deletion of KAP114, led to specific mislocalization of Sua7p to the cytoplasm. An interaction between Sua7p and TBP was also detected in cytosol raising the possibility that both Sua7p and TBP can be coimported by Kap114p. We have also shown that Kap114p possesses multiple overlapping binding sites for its partners, Sua7p, Nap1p and H2A and H2B, as well as RanGTP and nucleoporins. In addition, we have assembled an in vitro complex containing Sua7p, Nap1p, and histones H2A and H2B, suggesting that this Kap may import several proteins simultaneously. The import of more than one cargo at a time would increase the efficiency of each import cycle and may allow the regulation of coimported cargoes.


{dagger}These authors contributed equally to this study.

{sect}Present address: Division of Pulmonary and Critical Care Medicine, School of Medicine, Johns Hopkins University, Baltimore, MD 21287.

Address correspondence to: Lucy F. Pemberton (lfp2n{at}virginia.edu)




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