Isolation and Characterization of A Novel Human Radio-Susceptibility Gene, hNp95

• Project/Area Number: 15310044
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: National Institute of Radiological sciences
• Principal Investigator: FUJIMORI Akira (2004) National Institute of Radiological sciences, Research Center for Radiation Safety, Researcher, 放射線安全研究センター, 研究員 (50314183); 巽 紘一 (2003) 独立行政法人放射線医学総合研究所, 放射線安全研究センター, 研究員 (30131022)
• Co-Investigator(Kenkyū-buntansha): TATSUMI Kouichi National Institute of Radiological sciences, Research Center for Radiation Safety, Researcher, 放射線安全研究センター, 研究員 (30131022) ; MUTO Masahiro National Institute of Radiological sciences, Research Center for Radiation Safety, Researcher, 放射線安全研究センター, 研究員 (80166230) ; TAHAHAGI Masahiko National Institute of Radiological sciences, Research Center for Radiation Safety, Researcher, 放射線安全研究センター, 研究員 (30260235) ; KUBO Eiko National Institute of Radiological sciences, Research Center for Radiation Safety, Researcher, 放射線安全研究センター, 研究員 (40161658) ; 藤森 亮 独立行政法人放射線医学総合研究所, 放射線安全研究センター, 研究員 (50314183)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥5,500,000 (Direct Cost: ¥5,500,000); Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000); Fiscal Year 2003: ¥3,400,000 (Direct Cost: ¥3,400,000)
• Keywords: DNA binding protein / DNA damage / nuclear protein / DNA replication / 一本鎖DNA / MAR DNA / ゲルシフト / 蛋白質-DNA相互作用 / 高次DNA構造 / クロマチン / DNA複製点 / PCNA / hydroxyurea / アンチセンスcDNA / X線感受性 / 紫外線 / MNNG / トポイソメラーゼII α

Research Abstract

Murine nuclear protein NP95 has been shown to underlie resistance to ionizing radiation and other DNA insults or replication arrests in embryonic stem (ES) cells. Using the databases for expressed sequenced tags and a two-step PCR procedure, we isolated hNp95, the full-length human homologue of the murine NP95 cDNA, with a single open reading frame (ORF) encoding a polypeptide of 793 amino acids of 73.3% homology to NP95. The ORF of the hNP95 cDNA is identical to UHRF1 and only differs by two amino acids and seven nucleotides from that of the inverted CCAAT box binding protein of 90 kDa (ICBP90) that has been reported to regulate the transcription of the topoisomerase II gene. The hNp95 gene, assigned to 19p13.3, consists of 18 exons, spanning 60 kb, whereas the ICBP90 gene has been reported to comprise 6 exons. Several stable transformants from HEK293 and WI-38 cells that had been transfected with the antisense hNP95 cDNA were more sensitive to X-rays, UV-light and hydroxyurea than corresponding parental cells. hNP95 loss did not affect the activities of topoisomerase II□in HEK293 cells. These findings collectively indicate that the hNp95 gene is the human functional orthologue of the murine Np95 gene.

Research Products

• [Journal Article] Isolation and Characterization of A Novel Human Radio-Susceptibility Gene, hNp95 (2006)
  • Author(s): Masahiro Muto, Akira Fujimori, Mituru Nenoi, Kazuhiro Daino, Yoichi Matsuda, Asato Kuroiwa, Eiko Kubo, Yasuyoshi Kanari, Makoto Utsuno, Hideo Tsuji, Hideki Ukai, Kazuei Mita, Kouichi Tatsumi
  • Journal Title: Radiation Research (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] 抗ヒトhNP95モノクローナル抗体 (2003)
  • Inventor(s): 武藤正弘, 巽紘一
  • Industrial Property Rights Holder: 独立行政法人 放射線医学総合研究所
  • Patent Publication Number: 2005-087120
  • Filing Date: 2003-09-18
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Furuno-Fukushi, I.: "Effect of Atm Disruption on Spontaneously Arising and Radiation-Induced Deletion Mutations in Mouse Liver."Radiation Research. 160. 549-558 (2003)
• [Publications] Fukuchi, K.: "The Association of Cyclin A and Cyclin Kinase Inhibitor P21 in Response to Gamma-Irradiation Requires the Cdk2 Binding Region, but Not the Cy Motif."Biochimica et Biophysica Acta. 1642. 163-171 (2003)
• [Publications] Lenarczyk, M.: "The "Pro-Drug" Ribcys Decreases the Mutagenicity of High-Let Radiation in Cultured Mammalian Cell."Radiation Research. 160. 579-583 (2003)
• [Publications] Fukumura, R.: "A Sensitive Transcriptome Analysis Method That Can Detect Unknown Transcripts A Sensitive Transcriptome Analysis Method That Can Detect Unknown Transcripts."Nucleic Acids Research. 31. e94 (2003)
• [Publications] Watanabe, F.: "Involvement of DNA-Dependent Protein Kinase in Down-Regulation of Cell Cycle Progression."The International Journal of Biochemistry & Cell Biology. 35. 432-440 (2003)