Acrylamide catalytically inhibits topoisomerase II in V79 cells.
The vinyl monomer acrylamide is characterized by the presence of an alpha,beta-unsaturated carbonyl group that makes it reactive towards thiol, hydroxyl or amino groups and towards the nucleophilic centers in DNA. The ability of acrylamide to chemically modify protein thiols has prompted us to consider topoisomerase II as one possible target of acrylamide, since agents targeting protein sulfhydryl groups act as either catalytic inhibitors or poisons of topoisomerase II. Nuclear extracts from V79 Chinese hamster cells incubated with acrylamide reduced topoisomerase II activity as inferred by an inability to convert kinetoplast DNA to the decatenated form. Nuclear extracts incubated with acrylamide pre-incubated with DTT converted kinetoplast DNA to the decatenated form, suggesting that acrylamide influences topoisomerase II activity through reaction with sulfhydryl groups on the enzyme. Furthermore, acrylamide did not induce the pBR322 DNA cleavage, as assessed by cleavage assay; thus, it cannot be regarded as a poison of topoisomerase II. As a catalytic inhibitor, acrylamide antagonizes the effect of etoposide, a topoisomerase II poison, as determined by clonogenic assay in V79 cells. This antagonism is confirmed by band depletion assay, from which it can be inferred that acrylamide reduces the level of catalytically active cellular topoisomerase II available for the action of etoposide.
Sciandrello
Giulia
G
Dipartimento di Biologia Cellulare e dello Sviluppo A Monroy, Università di Palermo, Viale delle Scienze, Parco d'Orleans, 90128 Palermo, Italy. giscian@unipa.it
Mauro
Maurizio
M
Caradonna
Fabio
F
Catanzaro
Irene
I
Saverini
Marghereth
M
Barbata
Giusi
G
D020106:Acrylamide / Q000494:pharmacology*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D002460:Cell Line; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure; D003114:Colony-Forming Units Assay; D006224:Cricetinae; D053837:DNA Cleavage / Q000187:drug effects; D018105:DNA, Kinetoplast / Q000235:genetics / Q000378:metabolism; D005047:Etoposide / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D059005:Topoisomerase II Inhibitors
D000972:Antineoplastic Agents, Phytogenic; D018105:DNA, Kinetoplast; D019384:Nucleic Acid Synthesis Inhibitors; D059005:Topoisomerase II Inhibitors; D020106:Acrylamide; D005047:Etoposide