Human small cell lung cancer NYH cells resistant to the bisdioxopiperazine ICRF-187 exhibit a functional dominant Tyr165Ser mutation in the Walker A ATP binding site of topoisomerase II alpha

FEBS Lett. 2002 Jun 5;520(1-3):161-6. doi: 10.1016/s0014-5793(02)02805-3.

Abstract

Bisdioxopiperazine anti-cancer agents are catalytic inhibitors of topoisomerase II which by unknown means lock the enzyme in a closed clamp form and inhibit its ATPase activity. In order to demarcate a putative pharmacophore, we here describe a novel Tyr165Ser mutation in the enzyme's Walker A ATP binding site leading to specific bisdioxopiperazine resistance when transformed into a temperature-conditional yeast system. The Tyr165Ser mutation differed from a previously described Arg162Gln by being heterozygous and by purified Tyr165Ser enzyme being drug-resistant in a kinetoplast DNA decatenation enzymatic assay. This suggested dominant nature of Tyr165Ser was supported by co-transformation studies in yeast of plasmids carrying wild type and mutant genes. These results enable a model of the bisdioxopiperazine pharmacophore using the proposed asymmetric ATP hydrolysis of the enzyme.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Amino Acid Substitution
  • Antigens, Neoplasm
  • Antineoplastic Agents / pharmacology*
  • Binding Sites / genetics
  • Carcinoma, Small Cell / enzymology
  • Carcinoma, Small Cell / genetics*
  • Carcinoma, Small Cell / pathology
  • Cell Division / drug effects
  • Cell Division / genetics
  • DNA Topoisomerases, Type II / genetics*
  • DNA Topoisomerases, Type II / metabolism
  • DNA-Binding Proteins
  • Dose-Response Relationship, Drug
  • Drug Resistance, Fungal / genetics
  • Drug Resistance, Neoplasm / genetics
  • Genotype
  • Humans
  • Lung Neoplasms / enzymology
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology
  • Mutation
  • Protein Binding
  • Razoxane / pharmacology*
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Transformation, Genetic
  • Tumor Cells, Cultured / drug effects

Substances

  • Antigens, Neoplasm
  • Antineoplastic Agents
  • DNA-Binding Proteins
  • Razoxane
  • Adenosine Triphosphate
  • DNA Topoisomerases, Type II