Nuclear function and DNA topoisomerases

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(1) Epitope analysis of anti-DNA topoisomerase II monoclonal antibodyWe established over 100 hybridoma cells secreting anti-DNA topoisomerase II (topo II). We selected some and determined each epitope of oligopeptide level. One of them (6H8) can recognize topo II of every eucaryotes and another (8D2) is specific to proliferating cells and useful for diagnosis for neoplastic cells.(2) Analysis of topoisomerase of filamentous fungi and yeast.We cloned tope II gene of Aspergillus, Penicillium and Candida and analyzed nuclear localization signals at C-terminus region by Ala scanning method. From their sequence data we developed rapid PCR analysis for diagnosis test.(3) Novel site of localization of human topo II in nuclear regionIn addition to nuclear localization signal of human topo IIβ, we detected a novel signal sequence for the distinct localization site close to nucleolus. This signal is also found in several proteins such as RNA binding proteins and NPI46 (nucleolar peptidyl prolyl cis trans isomerase).(4) The monoclonal antibodies against human DNA topoisomerase IIαextensively used and characterized throughout this research project was filed for patent application by Mitsubishi-Kagaku and now comercialized for the diagnostic reagent of malignant tumor cells.