Adozelesin triggers DNA damage response pathways and arrests SV40 DNA replication through replication protein A inactivation.
The cyclopropylpyrroloindole anti-cancer drug, adozelesin, binds to and alkylates DNA. Treatment of human cells with low levels of adozelesin results in potent inhibition of both cellular and simian virus 40 (SV40) DNA replication. Extracts were prepared from adozelesin-treated cells and shown to be deficient in their ability to support SV40 DNA replication in vitro. This effect on in vitro DNA replication was dependent on both the concentration of adozelesin used and the time of treatment but was not due to the presence of adozelesin in the in vitro assay. Adozelesin treatment of cells was shown to result in the following: induction of p53 protein levels, hyperphosphorylation of replication protein A (RPA), and disruption of the p53-RPA complex (but not disruption of the RPA-cdc2 complex), indicating that adozelesin treatment triggers cellular DNA damage response pathways. Interestingly, in vitro DNA replication could be rescued in extracts from adozelesin-treated cells by the addition of exogenous RPA. Therefore, whereas adozelesin and other anti-cancer therapeutics trigger common DNA damage response markers, adozelesin causes DNA replication arrest through a unique mechanism. The S phase checkpoint response triggered by adozelesin acts by inactivating RPA in some function essential for SV40 DNA replication.
Liu
J S
JS
Department of Microbiology and the Center for Microbial Pathogenesis, State University of New York School of Medicine and Biomedical Sciences, Buffalo, New York 14214, USA.
Kuo
S R
SR
McHugh
M M
MM
Beerman
T A
TA
Melendy
T
T
D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D016590:Aphidicolin / Q000494:pharmacology; D001572:Benzofurans; D002461:Cell Line, Transformed; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology*; D053138:Cyclohexenes; D004261:DNA Replication / Q000187:drug effects*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D000080890:Duocarmycins; D006801:Humans; D007211:Indoles; D007700:Kinetics; D010766:Phosphorylation; D051716:Replication Protein A; D016196:S Phase; D013539:Simian virus 40 / Q000187:drug effects / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D018906:Antineoplastic Agents, Alkylating; D001572:Benzofurans; D003509:Cyclohexanecarboxylic Acids; D053138:Cyclohexenes; D004268:DNA-Binding Proteins; D000080890:Duocarmycins; D007211:Indoles; C497167:RPA1 protein, human; D051716:Replication Protein A; D016159:Tumor Suppressor Protein p53; D016590:Aphidicolin; C060090:adozelesin