Interaction between Pur α and ssCRE, and Pur α binding proteins

• Project/Area Number: 14580741
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Osaka University
• Principal Investigator: KUO Che-hui KUO,Che-hui, 医学系研究科, 助教授 (50126570)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000); Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: Purα / PUR element / Overlay assay / Gel shift assay / Brain / Nuclear Extract / Calmodulin / Single stranded DNA (ssDNA) / 構造変化 / 組織化学 / 転写 / 複製 / 神経

Research Abstract

Purα is an abundant protein in the brain and binds to a (GGN)n sequence, PUR element. The increased-immunoreactivity or silver staining of GST-Pur α in the presence of PUR element (ssCRE: single stranded cAMP response element) with or without CaM reflects well the DNA-binding activity of Purα. The similar results were observed with endogenous Purα from mouse brain. These data were just comparable to those of the gel shift assay and sensitivity with the immunoblot was comparable with that of the gel shift assay.
We also tried to search for Purα binding proteins (PurBPs) by the overlay assay with GSTPurα as a ligand. Three PurBPs were found mostly in the nuclear extract (N.Ext.) and they w6re not detected by the pretreatment of N.Ext. with trypsin, but not with RNase or DNase. The three PurBPs disappeared by the addition of ssCRE containing a PUR element, but not by ΔGGN ssCRE). The PurBPs were abundantly expressed in the brain as Purα.

Research Products

• [Publications] Ling-Hui Zeng, Takahiro Fujimoto, Naomasa Miki, Che-Hui Kuo, et al.: "Characterization of novel Purα-binding proteins in mouse brain"Neurochemistry International. (in press). (2004)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] LingHui Zeng, Takahiro Fujimoto, Emi Kumamaru, Yasuyuki Irie, Naomasa, Miki Che-Hul Kuol: Characterization of novel Purα binding proteins in mouse brain. Neurochemistry International. (in press). (2004)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ling-Hui Zeng, Takahiro Fujimoto, Naomasa Miki, Che-Hui Kuo et al.: "Characterization of novel Purα-binding proteins in mouse brain"Neurochemistry International. (in press). (2004)