Characterization of PSKH1, a novel human protein serine kinase with centrosomal, golgi, and nuclear localization.
We report here the characterization of PSKH1, a novel human protein serine kinase with multiple intracellular localizations. The gene consists of three exons distributed over 35 kb of genomic DNA in region 16q22.1. The 3.4-kb cDNA predicts a protein of 424 amino acids with a calculated molecular mass of 48.1 kDa and pI of 9.6. PSKH1 is expressed in all tissues and cell lines tested as shown by Northern blots, with the highest level of abundance in testis. PSKH1 displays the highest level of similarity with rat CaM kinase I (50. 2%) over 259 amino acids in the conserved catalytic region, but lacks significant homology with proteins in the database outside the catalytic core. Polyclonal antibodies have been raised, and indirect immunofluorescence microscopy of untransfected COS-1 cells suggests that PSKH1 is localized in the Brefeldin A-sensitive Golgi compartment, at centrosomes, in the nucleus with a somewhat speckle-like presence, and more diffusely in the cytoplasm. The presence in the centrosome appears to be enhanced during osmotic stress. Immunoisolated PSKH1 does not phosphorylate any of the common kinase substrates in vitro, but autophosphorylates exclusively serines within its COOH-terminal region in an intermolecular fashion. Furthermore, autophosphorylation activity is repressed upon addition of Ca(2+)/CaM, suggesting that PSKH1 activity depends on Ca(2+) concentration in vivo.
Brede
G
G
Biotechnology Center of Oslo, University of Oslo, Gaustadalleen 21, Oslo, 0349, Norway.
Solheim
J
J
Tröen
G
G
Prydz
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000201:enzymology*; D018385:Centrosome / Q000201:enzymology*; D003001:Cloning, Molecular; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D021382:Protein Sorting Signals; D021381:Protein Transport; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000378:metabolism; D013737:Testis / Q000201:enzymology; D014018:Tissue Distribution
D009687:Nuclear Proteins; D021382:Protein Sorting Signals; D012694:Serine; C417548:PSKH1 protein, human; D017346:Protein Serine-Threonine Kinases; C517596:CAMK1 protein, human; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C517599:Camk1 protein, rat; C517682:Pnck protein, rat