Nuclear function and DNA topoisomerases

核功能和 DNA 拓扑异构酶

• 批准号: 09558095
• 负责人: KIKUCHI Akihiko
• 金额: $ 3.33万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09558095/
• 关键词: DNA topoisomerase; Monoclonal Antibody; Epitopes; Cell Division Cycle; Signal sequence; Nuclear localization; Diagnosis and therapy; Mycology; 機能領域; たんぱく分解; 細胞内局在化; 酵母; DNAポイソメラーゼ; 活性阻害; 核移行シグナル; 種特異性; クリーバブル・コンプレックス; 大腸菌による発現; トポIIα・トポI

(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.

(1)抗DNA拓扑异构酶II单抗的表位分析我们建立了100多株分泌抗DNA拓扑异构酶II的杂交瘤细胞。我们选择了一些，并测定了每个表位的寡肽水平。(2)丝状真菌和酵母菌的拓扑异构酶分析。我们克隆了曲霉、青霉和念珠菌的拓扑酶II基因，并用ALA扫描法分析了C-末端的核定位信号。根据它们的序列数据，我们建立了用于诊断试验的快速聚合酶链式反应分析。(3)人Topo II在核区的新定位位点除了人Topo IIβ的核定位信号外，我们还检测到了靠近核仁的不同定位位点的新信号序列。该信号在核糖核酸结合蛋白和NPI46(核仁肽基脯氨酰顺式反式异构酶)等蛋白质中也可发现。(4)在整个研究项目中广泛使用和表征的抗人α拓扑异构酶II的单抗已被三菱香谷申请专利，现已商品化，用于恶性肿瘤细胞的诊断试剂。

项目成果

K.MATSUMOTO et al.: "Sperm chromatin decondensation by template activating factor I"Mol.Cell.Biol.. 19. 6940-6952 (1999)

K.MATSUMOTO 等人：“通过模板激活因子 I 进行精子染色质去浓缩”Mol.Cell.Biol.. 19. 6940-6952 (1999)

S.KATO and A.KIKUCHI: "“DNA Topoisomerase:The key enzyme that regulates DNA super structure" Nagoya Journal of Medical Sciences. 61. 11-26 (1998)

S.KATO 和 A.KIKUCHI：“DNA 拓扑异构酶：调节 DNA 超结构的关键酶”名古屋医学科学杂志 61. 11-26 (1998)。

K.-H. KIM et al.: "Cloning and characterization of the gene encoding Aspergillus nidulans"GENE. 236. 293-301 (1999)

K.-H。

Y.OKADA,Y.ITO,A.KIKUCHI,Y.NIMURA,S.YOSHIDA and M.SUZUKI: "Assignment of functional aminoacids around the active site of human DNA topoisomerase IIα"J.Biol.Chem.. 275,. 24630-24638 (2000)

Y.OKADA、Y.ITO、A.KIKUCHI、Y.NIMURA、S.YOSHIDA 和 M.SUZUKI：“人类 DNA 拓扑异构酶 IIα 活性位点周围的功能氨基酸的分配”J.Biol.Chem.. 275，。 -24638 (2000)

K.SUGIMOTO et al.: "Temporal and spatial distribution of DNA topoisomerase II alters during proliferation,differenciation and apoptosis in HL-60 cells" Blood. 91. 1407-1417 (1998)

K.SUGIMOTO 等人：“HL-60 细胞增殖、分化和凋亡过程中 DNA 拓扑异构酶 II 的时间和空间分布发生变化”血液。