Identification of proteins that interact with the central coiled-coil region of the human protein kinase NEK1.
NEK protein kinases are evolutionarily conserved kinases structurally related to the Aspergillus nidulans mitotic regulator NIMA. At least nine members of the NEK family in vertebrates have been described to date, but for most of them the interacting protein partners are unknown. The pleiotropic deleterious effects and the formation of kidney cysts caused by NEK1 mutation in mice emphasize its involvement in the regulation of diverse cellular processes and in the etiology of polycystic kidney disease (PKD), respectively. Here we report the identification of proteins that interacted with the human NEK1 protein kinase in a yeast two-hybrid screen of a human fetal brain cDNA library, using the catalytic and regulatory domains of NEK1 separately as baits. These proteins are known to take part either in the development of PKD, in the double-strand DNA break repair at the G2/M transition phase of the cell cycle, or in neural cell development. The proteins involved in PKD include the motor protein KIF3A and the proteins tuberin and alpha-catulin. Mapping studies of the human NEK1 regulatory domain (NRD) indicated a strong interaction of most of the proteins retrieved from the library with putative coiled coils located in the central region of NRD. Our results give further support to the previous observation that NEK1 is of functional importance for the etiology of PKD.
Surpili
Marcelo J
MJ
Centro de Biologia Molecular Estrutural (CEBIME), Laboratório Nacional de Luz Síncrotron (LNLS), Rua Giuseppe Máximo Scolfaro 10.000, CP 6192, 13084-971 Campinas, SP, Brazil.
Delben
Tatiana M
TM
Kobarg
Jörg
J
D000818:Animals; D001923:Brain Chemistry; D018797:Cell Cycle Proteins; D005333:Fetus; D006801:Humans; D016547:Kinesins / Q000302:isolation & purification; D051379:Mice; D000072380:NIMA-Related Kinase 1; D019151:Peptide Library; D010766:Phosphorylation; D007690:Polycystic Kidney Diseases / Q000201:enzymology / Q000473:pathology; D025941:Protein Interaction Mapping / Q000379:methods; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*; D012097:Repressor Proteins / Q000302:isolation & purification; D016415:Sequence Alignment; D013379:Substrate Specificity; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; D020798:Two-Hybrid System Techniques
D018797:Cell Cycle Proteins; C087415:KIF3A protein, human; C490202:Kif3a protein, mouse; D019151:Peptide Library; D012097:Repressor Proteins; C000624653:TSC2 protein, human; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; D011505:Protein-Tyrosine Kinases; C000606914:NEK1 protein, human; D000072380:NIMA-Related Kinase 1; C000606920:Nek1 protein, mouse; D017346:Protein Serine-Threonine Kinases; D016547:Kinesins