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© The Rockefeller University Press, 0021-9525/1998//1447 $5.00
The Journal of Cell Biology, Volume 143, Number 6, , 1998 1447-1455


Article

A Novel Nuclear Import Pathway for the Transcription Factor TFIIS



Markus Albertini, Lucy F. Pemberton, Jonathan S. Rosenblum, and Günter Blobel

Laboratory of Cell Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, New York 10021

We have identified a novel pathway for protein import into the nucleus. We have shown that the previously identified but uncharacterized yeast protein Nmd5p functions as a karyopherin. It was therefore designated Kap119p (karyopherin with Mr of 119 kD). We localized Kap119p to both the nucleus and the cytoplasm. We identified the transcription elongation factor TFIIS as its major cognate import substrate. The cytoplasmic Kap119p exists as an approximately stoichiometric complex with TFIIS. RanGTP, not RanGDP, dissociated the isolated Kap119p/TFIIS complex and bound to Kap119p. Kap119p also bound directly to a number of peptide repeat containing nucleoporins in overlay assays. In wild-type cells, TFIIS was primarily localized to the nucleus. In a strain where KAP119 has been deleted, TFIIS was mislocalized to the cytoplasm indicating that TFIIS is imported into the nucleus by Kap119p. The transport of various substrates that use other karyopherin-mediated import or export pathways was not affected in a kap119{Delta} strain. Hence Kap119p is a novel karyopherin that is responsible for the import of the transcription elongation factor TFIIS.

Key Words: yeast • nucleus • import • karyopherin • transcription factor



Abbreviations used in this paper: DAPI, 4',6-diamidino-2-phenylindole; GFP, green fluorescent protein; Kap, karyopherin; NES, nuclear export sequence; NLS, nuclear localization sequence; NPC, nuclear pore complex; Nups, nucleoporins; PrA, protein A from S. aureus.

M. Albertini was supported by a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft (A1425/2-1) and J.S. Rosenblum from the National Institutes of Health (GM-18720).



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