title: A novel lysine-rich domain and GTP binding motifs regulate the nucleolar retention of human guanine nucleotide binding protein, GNL3L.
abstract: A variety of G-proteins and GTPases are known to be involved in nucleolar function. We describe here a new evolutionarily conserved putative human GTPase, guanine nucleotide binding protein-like 3-like (GNL3L). Genes encoding proteins related to GNL3L are present in bacteria and yeast to metazoa and suggests its critical role in development. Conserved domain search analysis revealed that the GNL3L contains a circularly permuted G-motif described by a G5-G4-G1-G2-G3 pattern similar to the HSR1/MMR1 GTP-binding protein subfamily. Highly conserved and critical residues were identified from a three-dimensional structural model obtained for GNL3L using the crystal structure of an Ylqf GTPase from Bacillus subtilis. We demonstrate here that GNL3L is transported into the nucleolus by a novel lysine-rich nucleolar localization signal (NoLS) residing within 1-50 amino acid residues. NoLS identified here is necessary and sufficient to target the heterologous proteins to the nucleolus. We show for the first time that the lysine-rich targeting signal interacts with the nuclear transport receptor, importin-beta and transports GNL3L into the nucleolus. Interestingly, depletion of intracellular GTP blocks GNL3L accumulation into the nucleolar compartment. Furthermore, mutations within the G-domains alter the GTP binding ability of GNL3L and abrogate wild-type nucleolar retention even in the presence of functional NoLS, suggesting that the efficient nucleolar retention of GNL3L involves activities of both basic NoLS and GTP-binding domains. Collectively, these data suggest that GNL3L is composed of distinct modules, each of which plays a specific role in molecular interactions for its nucleolar retention and subsequent function(s) within the nucleolus.
authors.affiliation: Laboratory of Molecular Virology, Centre for DNA Fingerprinting and Diagnostics, ECIL Road, Nacharam, Hyderabad 500076, India.
authors.forename: M R K Subba
authors.initials: MR
authors.lastname: Rao
authors.forename: Gita
authors.initials: G
authors.lastname: Kumari
authors.forename: D
authors.initials: D
authors.lastname: Balasundaram
authors.forename: R
authors.initials: R
authors.lastname: Sankaranarayanan
authors.forename: S
authors.initials: S
authors.lastname: Mahalingam
mesh_terms: D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D002466:Cell Nucleolus / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D008239:Lysine; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D028961:beta Karyopherins
chemical_list: C509155:GNL3L protein, human; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D028961:beta Karyopherins; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D008239:Lysine