title: Cyclin H is targeted to the nucleus by C-terminal nuclear localization sequences.
abstract: Cdk-activating kinase (CAK) is a trimeric complex consisting of cdk7, cyclin H, and MAT1, which activates the cell-cycle-regulating cdks through T loop phosphorylation. In addition, other substrates of the CAK complex have been identified when CAK is assembled with the TFIIH core proteins, thereby regulating transcription and nucleotide excision repair. Little is known about the regulation of the CAK complex through cyclin H. In this study we further analyzed cyclin H regulation and identified two basic clusters in the C terminus of the protein as putative nuclear localization sequences (NLSs). Fusion constructs of full-length and truncated cyclin H sequences demonstrated the functionality of the NLSs. A peptide-binding assay revealed that at least one NLS interacts with the nuclear import receptors importin alpha/beta. Phosphorylation in the vicinity of the NLSs by cyclin C/cdk8 or protein kinase CK2, however, does not influence the nuclear translocation of cyclin H.
authors.affiliation: Department of Biophysics, University of the Saarland, Building 76, 66421 Homburg/Saar, Germany. andrea.krempler@uniklinik-saarland.de
authors.forename: A
authors.initials: A
authors.lastname: Krempler
authors.forename: S
authors.initials: S
authors.lastname: Kartarius
authors.forename: J
authors.initials: J
authors.lastname: Günther
authors.forename: M
authors.initials: M
authors.lastname: Montenarh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D047390:Casein Kinase II / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D056745:Cyclin C; D056748:Cyclin H; D016213:Cyclins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D028901:alpha Karyopherins / Q000378:metabolism*; D028961:beta Karyopherins / Q000378:metabolism*
chemical_list: C503674:CCNC protein, human; C534751:CCNH protein, human; C503675:Ccnc protein, mouse; C534752:Ccnh protein, mouse; D056745:Cyclin C; D056748:Cyclin H; D016213:Cyclins; D019913:Nuclear Localization Signals; D028901:alpha Karyopherins; D028961:beta Karyopherins; C485184:enhanced green fluorescent protein; C087978:karyopherin alpha 2; D049452:Green Fluorescent Proteins; D047390:Casein Kinase II