A high through-put screen for small molecules modulating mcm2 phosphorylation identifies ryuvidine as an inducer of the dna damage response
View/ Open
Full Text
Date
2014-06-05Author
FitzGerald, Jennifer
Murillo, Laura S.
O'Brien, Gemma
O'Connell, Enda
O'Connor, Aisling
Wu, Kevin
Wang, Guan-Nan
Rainey, Michael D.
Natoni, Alessandro
Healy, Sandra
O'Dwyer, Michael
Santocanale, Corrado
Metadata
Show full item recordUsage
This item's downloads: 0 (view details)
Cited 7 times in Scopus (view citations)
Recommended Citation
FitzGerald, Jennifer; Murillo, Laura S. O'Brien, Gemma; O'Connell, Enda; O'Connor, Aisling; Wu, Kevin; Wang, Guan-Nan; Rainey, Michael D.; Natoni, Alessandro; Healy, Sandra; O'Dwyer, Michael; Santocanale, Corrado (2014). A high through-put screen for small molecules modulating mcm2 phosphorylation identifies ryuvidine as an inducer of the dna damage response. PLoS ONE 9 (6),
Published Version
Abstract
DNA replication is an essential process for cell division and as such it is a process that is directly targeted by several anticancer drugs. CDC7 plays an essential role in the activation of replication origins and has recently been proposed as a novel target for drug discovery. The MCM DNA helicase complex (MCM2-7) is a key target of the CDC7 kinase, and MCM phosphorylation status at specific sites is a reliable biomarker of CDC7 cellular activity. In this work we describe a cell-based assay that utilizes the "In Cell Western Technique'' (ICW) to identify compounds that affect cellular CDC7 activity. By screening a library of approved drugs and kinase inhibitors we found several compounds that can affect CDC7-dependent phosphorylation of MCM2 in HeLa cells. Among these, Mitoxantrone, a topoisomerase inhibitor, and Ryuvidine, previously described as a CDK4 inhibitor, cause a reduction in phosphorylated MCM2 levels and a sudden blockade of DNA synthesis that is accompanied by an ATM-dependent checkpoint response. This study sheds light on the previously observed cytotoxity of Ryuvidine, strongly suggesting that it is related to its effect of causing DNA damage.