title: A proline-tyrosine nuclear localization signal (PY-NLS) is required for the nuclear import of fission yeast PAB2, but not of human PABPN1.
abstract: Nuclear poly(A)-binding proteins (PABPs) are evolutionarily conserved proteins that play key roles in eukaryotic gene expression. In the fission yeast Schizosaccharomyces pombe, the major nuclear PABP, Pab2, functions in the maturation of small nucleolar RNAs as well as in nuclear RNA decay. Despite knowledge about its nuclear functions, nothing is known about how Pab2 is imported into the nucleus. Here, we show that Pab2 contains a proline-tyrosine nuclear localization signal (PY-NLS) that is necessary and sufficient for its nuclear localization and function. Consistent with the role of karyopherin β2 (Kapβ2)-type receptors in the import of PY-NLS cargoes, we show that the fission yeast ortholog of human Kapβ2, Kap104, binds to recombinant Pab2 and is required for Pab2 nuclear localization. The absence of arginine methylation in a basic region N-terminal to the PY-core motif of Pab2 did not affect its nuclear localization. However, in the context of a sub-optimal PY-NLS, we found that Pab2 was more efficiently targeted to the nucleus in the absence of arginine methylation, suggesting that this modification can affect the import kinetics of a PY-NLS cargo. Although a sequence resembling a PY-NLS motif can be found in the human Pab2 ortholog, PABPN1, our results indicate that neither a functional PY-NLS nor Kapβ2 activity are required to promote entry of PABPN1 into the nucleus of human cells. Our findings describe the mechanism by which Pab2 is imported into the nucleus, providing the first example of a PY-NLS import system in fission yeast. In addition, this study suggests the existence of alternative or redundant nuclear import pathways for human PABPN1.
authors.affiliation: RNA Group, Department of Biochemistry, Université de Sherbrooke, Sherbrooke, QC, Canada.
authors.forename: Pierre-Luc
authors.initials: PL
authors.lastname: Mallet
authors.forename: François
authors.initials: F
authors.lastname: Bachand
mesh_terms: D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D001120:Arginine / Q000378:metabolism; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019913:Nuclear Localization Signals; D039122:Poly(A)-Binding Protein I / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D039103:Poly(A)-Binding Protein II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D039102:Poly(A)-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011392:Proline / Q000737:chemistry; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism*
chemical_list: C582315:Kap104 protein, S pombe; D019913:Nuclear Localization Signals; C576588:PABPN1 protein, human; C582314:Pab2 protein, S pombe; D039122:Poly(A)-Binding Protein I; D039103:Poly(A)-Binding Protein II; D039102:Poly(A)-Binding Proteins; D029702:Schizosaccharomyces pombe Proteins; D028961:beta Karyopherins; D014443:Tyrosine; D001120:Arginine; D011392:Proline