2003 Fiscal Year Final Research Report Summary

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

Research Project

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
Keywords	Purα / PUR element / Overlay assay / Gel shift assay / Brain / Nuclear Extract / Calmodulin / Single stranded DNA (ssDNA)
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.

(2 results)