Protein kinase CK2 interacts with Chk2 and phosphorylates Mre11 on serine 649.
The Mre11-Rad50-Nbs1 protein complex has been known to be involved in a variety of DNA metabolic events that involve DNA double-strand breaks (DSBs). The phosphorylation of Mre11 is increased in response to ionizing radiation, which suggests that phosphorylation of Mre11 may be an important regulatory mechanism of this complex. Mre11-phosphorylating kinase activities were observed in Chk2 immunoprecipitates and HeLa nuclear extracts. Through the tandem affinity tagging system and conventional chromatography, this kinase was purified and identified as protein kinase CK2. CK2 phosphorylates Mre11 in vitro. In vitro kinase assay with a series of truncated Mre11 proteins as substrates for CK2 and site-directed mutagenesis showed that serine 649 of Mre11 is mainly phosphorylated by CK2 in vitro. In vivo labeling and phosphopeptide mapping analysis revealed that this phosphorylation occurs in vivo. These data implicate CK2 as a potential upstream regulator of Mre11 function.
Kim
Seong-Tae
ST
Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, 300 Chunchundong, Jangangu, Suwon, Kyonggido 440-746, Republic of Korea. stkim@med.skku.ac.kr
D000595:Amino Acid Sequence; D047390:Casein Kinase II / Q000378:metabolism*; D064447:Checkpoint Kinase 2; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D000076228:MRE11 Homologue Protein; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*
D004268:DNA-Binding Proteins; C121766:MRE11 protein, human; D012694:Serine; D064447:Checkpoint Kinase 2; C578634:CHEK2 protein, human; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D000076228:MRE11 Homologue Protein