Molecular mechanisms of GRP78-involving process to repair UVC-damaged DNA

2006 Fiscal Year Final Research Report Summary

• Project/Area Number: 17510040
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: Chiba University
• Principal Investigator: KITA Kazuko Chiba University, Graduate School of Medicine, Lecturer, 大学院医学研究院, 講師 (80302545)
• Co-Investigator(Kenkyū-buntansha): SUZUKI Toshikazu Chiba University, Graduate School of Medicine, Research Associate, 大学院医学研究院, 助手 (70270527) ; ICHIMURA Yoshinob Chiba University, Graduate School of Medicine, Research Associate, 大学院医学研究院, 助手 (80400993)
• Project Period (FY): 2005 – 2006
• Keywords: human cells / UV / DNA damage / DNA repair / ER molecular chaperone / GRP78 / sensitivity to cell death

Research Abstract

There are only a few reports about the roles of the 78-kDa glucose-regulated protein (GRP78) in human cell response to DNA-damaging stresses. We previously found that GRP78 is involved in resistance to UVC-induced cell death in human cells. We here investigated molecular mechanisms of GRP78-mediated UVC resistance. To investigate whether GRP78 is involved in resistance to a DNA-damaging agent, UVC (principally 254 nm in wavelength), we established human cells with down-regulation of GRP78 by transfection of human RSa cells with antisense cDNA for GRP 78. Using the cells, we found the new findings as below: 1) In the antisense cDNA-transfected cells DNA synthesis activity of whole cell extracts to repair UVC-irradiated plasmids in vitro were remarkably decreased compared with those in the control cells. However, neither addition of recombinant GRP78 protein to the extracts of the former cells recovered the repair synthesis activity, nor depletion of GRP78 from the extracts of the latter cells by immunoprecipitation decreased the activity. 2) The amount of XPA protein was lower in the extracts from the antisene cDNA-transfected cells than from the control cells. 3) GRP78-GFP, expressed in the cultured human cells, localized at perinuclear region. However, the localization did not change remarkably after UVC irradiation. 4) The antisense cDNA-transfected cells showed higher sensitivity to cisplatin-induced cell death, but only slightly higher sensitivity to X-ray-induced cell death than the control cells.
The present results suggest that GRP78 plays a protective role against UVC-induced cell death possibly via nucleotide excision repair (NER), at least in the human RSa cells tested. Molecular mechanisms of the involvement of GRP78 in NER process remain unknown. However, these findings also suggest that GRP78 may not associate directly with repair complex involved the process, but stabilize the NER components or regulate the metabolism of the components in the nucleus.

Research Products

• [Journal Article] Life science of gravity-changing stress; modulation of UV- mutagenicity in human RSb cells by serum factors from volunteers who experienced parabolic flight. (2006)
  • Author(s): Suzuki, N., Chi, X., Takahashi, S., Nomura, J., Sugaya, S., Ichimura, Y., Zhai, L., Tong, X., Kita, K.
  • Journal Title: J. Int. Soc. Life Info. Sci. 24, 2 Pages: 362-363
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Crystallization and preliminary X-ray analysis of Atg3. (2006)
  • Author(s): Yamada Y, Suzuki NN, Fujioka Y, Ichimura Y, Ohsumi Y, Inagaki F
  • Journal Title: Acta. Crystallograph. Sect. F. Struct. Biol. Cryst. Commun. 62 Pages: 1016-1017
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Two newly identified sites in the ubiquitin-like protein Atg8 are essential for autophagy. (2006)
  • Author(s): Amar N, Lustig G, Ichimura Y, Ohsumi Y, Elazar Z
  • Journal Title: EMBO Rep. 7 Pages: 635-642
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Reduction of GRP78 expression leads to activation of endoplasmic reticulum (ER) stress response in HeLa cells. (2006)
  • Author(s): Suzuki T., Kites, K., Lu, J., Suzuki N.
  • Journal Title: Abstract of the 79^<th> Annual Meeting of the Japanese Biochemical Society Pages: 432
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Search for proteins interacted with HSP27 in UVC-resistant human cells. (2006)
  • Author(s): Kites K., Tong, X., Karata, K., Sugaya, S., Jin, Y., Sato, M., Tomonaga, T., Nomura.F., Suzuki.N.
  • Journal Title: Abstract of the 49^<th> Annual Meeting of the Japan Radiation Research Society Pages: 125
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Life science of gravity-changing stress ; modulaiton of UV-mutagenicity in human RSb cells by serum factors from volunteers who experienced parabolic flight. (2006)
  • Author(s): Suzuki, N., Chi, X., Takahashi, S., Nomura, J., Sugaya, S., Ichimura, Y., Zhai, L., Tong, X., Kita, K.
  • Journal Title: J.Int.Soc.Life Info.Sci. 24 Pages: 362-363
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Crystallization and preliminary X-ray analysis of Atg3. (2006)
  • Author(s): Yamada Y, Suzuki NN, Fujioka Y, Ichimura Y, Ohsumi Y, Inagaki F.
  • Journal Title: Acta.Crystallograph.Sect.F.Struct.Biol.Cryst.Commun. 62 Pages: 1016-1017
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two newly identified sites in the ubiquitin-like protein Atg8 are essential for autophagy. (2006)
  • Author(s): Amar N, Lustig G, Ichimura Y, Ohsumi Y, Elazar Z.
  • Journal Title: EMBO Rep. 7 Pages: 635-642
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Decreased cell survival and DNA repair capacity after UVC irradiation in association with down-regulation of GRP78/BiP in human RSa cells. (2005)
  • Author(s): Zhai, L., Kita, K., Wano, C., Wu, Y., Sugaya, S., Suzuki, N.
  • Journal Title: Exp. Cell Res. 305 Pages: 244-252
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Down-regulation of SMT3 A gene expression is association with DNA synthesis induction after X-ray irradiation in nevoid basal cell carcinoma syndrome (NBCCS) cells. (2005)
  • Author(s): Sugaya, S., Nakanishi, H., Tanzawa H., Sugita, K., Kita, K., Suzuki, N.
  • Journal Title: Mut. Res. 578 Pages: 327-332
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Enhanced expression of transferring receptor confers UV-resistance in human and monkey cells. (2005)
  • Author(s): Chen, Z., Nomura, J., Suzuki, T., Suzuki, N.
  • Journal Title: J. Radiat.Res. 46 Pages: 443-451
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Leupeptin-sensitive protease involved in cell survival after X-ray irradiation in human RSa cells. (2005)
  • Author(s): Zhang, H., Karata, K., Matsushita, K., Takahashi, S., Tong X., Lu, J., Zhu, C., Sugaya, S., Suzuki, T., Suzuki, N.
  • Journal Title: Cell Biol. Int. 29 Pages: 662-668
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Increased expression of GRP94 protein is associated with decreased sensitivity to X-rays in cervical cancer cel lines. (2005)
  • Author(s): Kubota, H., Suzuki, T., Lu, J., Takahashi, S., Sugaya, S., Sekiya, S., Suzuki, N
  • Journal Title: Int. F. Radiat. Biol. 81 Pages: 701-709
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of GRP78 in cell survival and DNA repair in UVC- and cisplatin-treated human cells. (2005)
  • Author(s): Kita, K., Zhai, L., Sugaya, S., Suzuki N.
  • Journal Title: Abstract of the 78^<th> Annual Meeting of the Japanese Biochemical Society 77 Pages: 952
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Decreased cell survival and DNA repair capacity after UVC irradiation in association with down-regulation of GRP78/BiP in human RSa cells. (2005)
  • Author(s): Zhai, L., Kita, K., Wano, C., Wu, Y., Sugaya, S., Suzuki, N.
  • Journal Title: Exp.Cell Res. 305 Pages: 244-252
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Down-regulation of SMT3A gene expression is association with DNA synthesis induction after X-ray irradiation in nevoid basal cell carcinoma syndrome (NBCCS) cells. (2005)
  • Author(s): Sugaya, S., Nakanishi, H., Tanzawa, H., Sugita, K., Kita, K., Suzuki, N.
  • Journal Title: Mut.Res. 578 Pages: 327-332
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Enhanced expression of transferring receptor confers UV-resistance in human and monkey cells. (2005)
  • Author(s): Chen, Z., Nomura, J., Suzuki, T., Suzuki, N.
  • Journal Title: J.Radiat.Res. 46 Pages: 443-451
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Leupeptin-sensitive protease involved in cell survival after X-ray irradiation in human RSa cells. (2005)
  • Author(s): Zhang, H., Karata, K., Matsushita, K., Takahashi, S., Tong, X., Lu, J., Zhu, C., Sugaya, S., Suzuki, T., Suzuki, N.
  • Journal Title: Cell Biol.Int. 29 Pages: 662-668
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Increased expression of GRP94 protein is associated with decreased sensitivity to X-rays in cervical cancer cell lines. (2005)
  • Author(s): Kubota, H., Suzuki, T., Lu, J., Takahashi, S., Sugaya, S., Sekiya, S., Suzuki, N.
  • Journal Title: Int.F.Radiat.Biol. 81 Pages: 701-709
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Modulation of mutability in UV-irradiated human RSb cells by serum obtained from parabolic flight volunteers.
  • Author(s): Chi, X, Takahashi, S., Nomura J., Sugaya S., Ishimura Y., Zhai L., Tong X., Kita K., Suzuki N.
  • Journal Title: J. Int. Soc. Life Info. Sci. (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Modulation of mutability in UV-irradiated human RSb cells by serum obtained from parabolic flight volunteers.
  • Author(s): Chi, X., Takahashi, S., Nomura, J., Sugaya, S., Ishimura, Y., Zhai, L., Tong, X., Kita, K., Suzuki, N.
  • Journal Title: J.Int.Soc.Life Info.Sci. (in press)
  • Description: 「研究成果報告書概要(欧文)」より