Identification of an unconventional nuclear localization signal in human ribosomal protein S2

• Published in: Biochemical and biophysical research communications Vol. 335; no. 1; pp. 146 - 153
• Main Authors: Antoine, M., Reimers, K., Wirz, W., Gressner, A.M., Müller, R., Kiefer, P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.09.2005
• Subjects: 60 APPLIED LIFE SCIENCES; Active Transport, Cell Nucleus; AMINO ACID SEQUENCE; AMINO ACIDS; Animals; beta-Galactosidase - genetics; beta-Galactosidase - metabolism; Cell Nucleolus - metabolism; Cell Nucleus - metabolism; Cercopithecus aethiops; CHIMERAS; COS Cells; CYTOPLASM; Cytoplasm - metabolism; GALACTOSIDASE; Humans; IMPORTS; Molecular Sequence Data; MUTANTS; Mutation - genetics; Nuclear localization sequence; Nuclear Localization Signals - physiology; NUCLEOLI; Nup153; Protein Binding; RECEPTORS; Ribosomal protein S2; Ribosomal Proteins - chemistry; Ribosomal Proteins - genetics; Ribosomal Proteins - metabolism; Sequence Alignment; Subcellular localization; VP22; Nup153; VP22; Nuclear localization sequence; Ribosomal protein S2; Subcellular localization
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2005.07.069

Ribosomal proteins must be imported into the nucleus after being synthesized in the cytoplasm. Since the rpS2 amino acid sequence does not contain a typical nuclear localization signal, we used deletion mutant analysis and rpS2–β-galactosidase chimeric proteins to identify the nuclear targeting domains in rpS2. Nuclear rpS2 is strictly localized in the nucleoplasm and is not targeted to the nucleoli. Subcellular localization analysis of deletion mutants of rpS2–β-galactosidase chimeras identified a central domain comprising 72 amino acids which is necessary and sufficient to target the chimeric β-galactosidase to the nucleus. The nuclear targeting domain shares no significant similarity to already characterized nuclear localization signals in ribosomal proteins or other nuclear proteins. Although a Nup153 fragment containing the importinβ binding site fused to VP22 blocks nuclear import of rpS2–β-galactosidase fusion proteins, nuclear uptake of rpS2 could be mediated by several import receptors since it binds to importinα/β and transportin.