title: Characterization of karyopherin cargoes reveals unique mechanisms of Kap121p-mediated nuclear import.
abstract: In yeast there are at least 14 members of the beta-karyopherin protein family that govern the movement of a diverse set of cargoes between the nucleus and cytoplasm. Knowledge of the cargoes carried by each karyopherin and insight into the mechanisms of transport are fundamental to understanding constitutive and regulated transport and elucidating how they impact normal cellular functions. Here, we have focused on the identification of nuclear import cargoes for the essential yeast beta-karyopherin, Kap121p. Using an overlay blot assay and coimmunopurification studies, we have identified 30 putative Kap121p cargoes. Among these were Nop1p and Sof1p, two essential trans-acting protein factors required at the early stages of ribosome biogenesis. Characterization of the Kap121p-Nop1p and Kap121p-Sof1p interactions demonstrated that, in addition to lysine-rich nuclear localization signals (NLSs), Kap121p recognizes a unique class of signals distinguished by the abundance of arginine and glycine residues and consequently termed rg-NLSs. Kap104p is also known to recognize rg-NLSs, and here we show that it compensates for the loss of Kap121p function. Sof1p is also transported by Kap121p; however, its import can be mediated by a piggyback mechanism with Nop1p bridging the interaction between Sof1p and Kap121p. Together, our data elucidate additional levels of complexity in these nuclear transport pathways.
authors.affiliation: Institute for Systems Biology, Seattle, Washington 98103, USA.
authors.forename: Deena M
authors.initials: DM
authors.lastname: Leslie
authors.forename: Wenzhu
authors.initials: W
authors.lastname: Zhang
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Timney
authors.forename: Brian T
authors.initials: BT
authors.lastname: Chait
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
authors.forename: Richard W
authors.initials: RW
authors.lastname: Wozniak
authors.forename: John D
authors.initials: JD
authors.lastname: Aitchison
mesh_terms: D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D028884:Karyopherins / Q000378:metabolism*; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D056489:Nucleoplasmins; D010750:Phosphoproteins / Q000378:metabolism; D025941:Protein Interaction Mapping; D021381:Protein Transport / Q000502:physiology*; D011506:Proteins / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D020718:Ribonucleoproteins, Small Nucleolar / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D015003:Yeasts / Q000235:genetics / Q000378:metabolism; D028961:beta Karyopherins
chemical_list: C101820:KAP104 protein, S cerevisiae; D028884:Karyopherins; C064793:NOP1 protein, S cerevisiae; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins; D011506:Proteins; D017411:Ribonucleoproteins, Small Nuclear; D020718:Ribonucleoproteins, Small Nucleolar; C081686:SOF1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D028961:beta Karyopherins