Waranun BuajeebXue ZhangHiroe OhyamaDavid HanRudee SuraritYong KimDavid T.W. WongMahidol UniversityChina Medical University ShenyangHarvard School of Dental MedicineUniversity of California, Los AngelesDavid Geffen School of Medicine at UCLAMolecular Biology Institute2018-07-242018-07-242004-03-19Biochemical and Biophysical Research Communications. Vol.315, No.4 (2004), 998-10030006291X2-s2.0-1342302367https://repository.li.mahidol.ac.th/handle/20.500.14594/21214Human DOC-1/CDK2AP1 gene encodes a growth suppressor protein of 12kDa (p12DOC-1/CDK2AP1). Recently, p12DOC-1/CDK2AP1has been shown to associate with cell cycle proteins including CDK2 and DNA polymerase α/primase. It negatively regulates CDK2 activities and suppresses DNA replication. Therefore, identification of other p12DOC-1/CDK2AP1interacting proteins might clarify its role in the cell cycle regulation and carcinogenesis. The purpose of this study was to identify additional p12DOC-1/CDK2AP1interacting proteins using the yeast two-hybrid system. Using human p12DOC-1/CDK2AP1as a bait in a liver cDNA library screening, cDNA clones identical to human DOC-1R transcript were identified. The interaction between p12DOC-1/CDK2AP1and p14DOC-1Rwas verified in vitro and in cells. GST pull-down assay and immunoprecipitation experiments confirmed the interaction between the two proteins. The critical region for p12DOC-1/CDK2AP1interaction with p14DOC-1Rwas defined to amino acids 20-25 by using a series of deletion mutants as baits in the yeast two-hybrid system. Our data indicated that p12DOC-1/CDK2AP1could associate with its homologous protein, p14DOC-1R. © 2004 Elsevier Inc. All rights reserved.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1016/j.bbrc.2004.02.003