Functional analysis of primary open-angle glaucoma associated WDR36 gene.

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 18591948
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Ophthalmology
• Research Institution: 独立行政法人国立病院機構(東京医療センター臨床研究センター)
• Principal Investigator: MINAMI Masayoshi 独立行政法人国立病院機構(東京医療センター臨床研究センター), Institution, department, title of position : Division of Molecular and Cellular Biology, National Institute of Sensory Organs, Postdoctoral fellow (70392800)
• Co-Investigator(Kenkyū-buntansha): IWATA Takeshi National Institute of Sensory Organs, Institution, department, title of position : Division of Molecular and Cellular Biology, Department manager (90374157)
• Project Period (FY): 2006 – 2007
• Keywords: gene / protein / neuroscience / WDR36

Research Abstract

It has been reported that four mutations of WD-repeat 36(WDR36) gene are involved in the onset of primary open-angle glaucoma in the United States. In this study, to analyze the function of WDR36, we intended to express and purify the recombinant protein, examined the localization of WDR36 protein, and made sure whether reported mutations are detected in Japanese population. When we introduced WDR36-expressed vector into E. coli., most of the recombinant proteins were insoluble. To improve an efficiency of solubility, we investigated the conditions of induction (such as induction temperature concentration of isopropyl-β-D-thiogalactoside and existence of molecular chaperone) . As a result, we were able to increase the solubility. We prepared genomic DNA from 70 Japanese glaucoma patients, amplified the reported mutations containing region by polymerase chain reaction, and determined the sequence. The patients specific mutations could not be detected. Therefore, we speculate the possibility that reported mutations are not associated with the onset of glaucoma. However, it is necessary to analyze more number of cases to confirm above results We carried out immunofluorescent staining experiments and revealed that the localization of wild type or mutant WDR36 protein in cultured cell lines (COS-1, NIH3T3 and RGC-5) is a cytosol. These results suggest that WDR36 protein may act not in a specific organelle but in a cytosol, and the insertion of mutations do not affect the distribution of molecule.

Research Products

• [Journal Article] HTRA1 promoter polymorphism predisposes Japanese to age-related macular degeneration. (2007)
  • Author(s): Yoshida T, DeWan A, Zhang H, Sakamoto R, Okamoto H, Minami M, Obazawa M, Mizota A, Tanaka M, Saito Y, Takagi I, Hoh J, Iwata T.
  • Journal Title: Molecular Vision 13 Pages: 545-548
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] HTRA1 promoter polymorphism predisposes Japanese to age-related macular degeneration. (2007)
  • Author(s): Yoshida T, DeWan A, Zhang H, Sakamoto R, Okamato H, Minami M, Obazawa M, Mizota A, Tanaka M, Saito Y, Takagi I, Hoh J, Iwata T.
  • Journal Title: Molesular Vision(perr-reviewed journal) 13 Pages: 545-548
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Interaction of secreted myocilin and the cell surface proteins on NIH3T3 cells. (2007)
  • Author(s): 皆見政好、岩田岳
  • Organizer: The Association for Research in Vision and Ophthalmology
  • Place of Presentation: アメリカ合衆国フロリダ州
  • Year and Date: 2007-05-05
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Interaction of secreted myocilin and the cell surface proteins on NIH3T3 cells. (2007)
  • Author(s): Minami M, Iwata T.
  • Organizer: The Association for Research in Vision Ophthalmology
  • Place of Presentation: FL, USA
  • Year and Date: 2007-05-05
  • Description: 「研究成果報告書概要(欧文)」より