The bipartite nuclear localization sequence of Rpn2 is required for nuclear import of proteasomal base complexes via karyopherin alphabeta and proteasome functions.
26 S proteasomes fulfill final steps in the ubiquitin-dependent degradation pathway by recognizing and hydrolyzing ubiquitylated proteins. As the 26 S proteasome mainly localizes to the nucleus in yeast, we addressed the question how this 2-MDa multisubunit complex is imported into the nucleus. 26 S proteasomes consist of a 20 S proteolytically active core and 19 S regulatory particles, the latter composed of two subcomplexes, namely the base and lid complexes. We have shown that 20 S core particles are translocated into the nucleus as inactive precursor complexes via the classic karyopherin alphabeta import pathway. Here, we provide evidence that nuclear import of base and lid complexes also depends on karyopherin alphabeta. Potential classic nuclear localization sequences (NLSs) of base subunits were analyzed. Rpn2 and Rpt2, a non-ATPase subunit and an ATPase subunit of the base complex, harbor functional NLSs. The Rpt2 NLS deletion yielded wild type localization. However, the deletion of the Rpn2 NLS resulted in improper nuclear proteasome localization and impaired proteasome function. Our data support the model by which nuclear 26 S proteasomes are assembled from subcomplexes imported by karyopherin alphabeta.
Wendler
Petra
P
Institut für Biochemie CCM, Charité, Universitätsmedizin Berlin, Monbijoustrasse 2, D-10117 Berlin, Germany.
Lehmann
Andrea
A
Janek
Katharina
K
Baumgart
Sabine
S
Enenkel
Cordula
C
D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D006868:Hydrolysis; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D028901:alpha Karyopherins / Q000378:metabolism*; D028961:beta Karyopherins / Q000378:metabolism*
D019913:Nuclear Localization Signals; C096166:RPN2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D028901:alpha Karyopherins; D028961:beta Karyopherins; D046988:Proteasome Endopeptidase Complex