Loss of CCDC6, the first identified RET partner gene, affects pH2AX S139 levels and accelerates mitotic entry upon DNA damage.
CCDC6 was originally identified in chimeric genes caused by chromosomal translocation involving the RET proto-oncogene in some thryoid tumors mostly upon ionizing radiation exposure. Recognised as a pro-apoptotic phosphoprotein that negatively regulates CREB1-dependent transcription, CCDC6 is an ATM substrate that is responsive to genotoxic stress. Here we report that following genotoxic stress, loss or inactivation of CCDC6 in cancers that carry the CCDC6 fusion, accelerates the dephosphorylation of pH2AX S139, resulting in defective G2 arrest and premature mitotic entry. Moreover, we show that CCDC6 depleted cells appear to repair DNA damaged in a shorter time compared to controls, based on reporter assays in cells. High-troughput proteomic screening predicted the interaction between the CCDC6 gene product and the catalytic subunit of Serin-Threonin Protein Phosphatase 4 (PP4c) recently identified as the evolutionarily conserved pH2AX S139 phosphatase that is activated upon DNA Damage. We describe the interaction between CCDC6 and PP4c and we report the modulation of PP4c enzymatic activity in CCDC6 depleted cells. We discuss the functional significance of CCDC6-PP4c interactions and hypothesize that CCDC6 may act in the DNA Damage Response by negatively modulating PP4c activity. Overall, our data suggest that in primary tumours the loss of CCDC6 function could influence genome stability and thereby contribute to carcinogenesis.
Istituto di Endocrinologia ed Oncologia Sperimentale, CNR, Naples, Italy.
Francesco
F
Merolla
Chiara
C
Luise
Mark T
MT
Muller
Roberto
R
Pacelli
Alfredo
A
Fusco
Angela
A
Celetti
D002460:Cell Line; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D004249:DNA Damage; D016195:G2 Phase; D020868:Gene Silencing; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008938:Mitosis; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D000090063:Proto-Oncogene Mas
C088832:CCDC6 protein, human; D003598:Cytoskeletal Proteins; C468783:H2AX protein, human; D006657:Histones; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D010749:Phosphoprotein Phosphatases; C080406:protein phosphatase 4