title: Mechanisms controlling subcellular localization of the G(1) cyclins Cln2p and Cln3p in budding yeast.
abstract: Different G(1) cyclins confer functional specificity to the cyclin-dependent kinase (Cdk) Cdc28p in budding yeast. The Cln3p G(1) cyclin is localized primarily to the nucleus, while Cln2p is localized primarily to the cytoplasm. Both binding to Cdc28p and Cdc28p-dependent phosphorylation in the C-terminal region of Cln2p are independently required for efficient nuclear depletion of Cln2p, suggesting that this process may be physiologically regulated. The accumulation of hypophosphorylated Cln2 in the nucleus is an energy-dependent process, but may not involve the RAN GTPase. Phosphorylation of Cln2p is inefficient in small newborn cells obtained by elutriation, and this lowered phosphorylation correlates with reduced Cln2p nuclear depletion in newborn cells. Thus, Cln2p may have a brief period of nuclear residence early in the cell cycle. In contrast, the nuclear localization pattern of Cln3p is not influenced by Cdk activity. Cln3p localization requires a bipartite nuclear localization signal (NLS) located at the C terminus of the protein. This sequence is required for nuclear localization of Cln3p and is sufficient to confer nuclear localization to green fluorescent protein in a RAN-dependent manner. Mislocalized Cln3p, lacking the NLS, is much less active in genetic assays specific for Cln3p, but more active in assays normally specific for Cln2p, consistent with the idea that Cln3p localization explains a significant part of Clnp functional specificity.
authors.affiliation: The Rockefeller University, New York, New York 10021, USA.
authors.forename: M E
authors.initials: ME
authors.lastname: Miller
authors.forename: F R
authors.initials: FR
authors.lastname: Cross
mesh_terms: D000595:Amino Acid Sequence; D016213:Cyclins / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D016193:G1 Phase; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D010766:Phosphorylation; D029701:Saccharomyces cerevisiae Proteins; D004718:Saccharomycetales / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C082721:CLN2 protein, S cerevisiae; C079895:CLN3 protein, S cerevisiae; D016213:Cyclins; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins