| Project/Area Number |
13672230
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Chemical pharmacy
|
|---|
| Research Institution | Teikyo University |
|---|
Principal Investigator |
KITTAKA Atsushi Teikyo University Faculty of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (00214833)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NOZOMI Saito Teikyo University Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助手 (80349258)
SUHARA Yoshitomo Kobe Pharmaceutical University Department of Hygienic Sciences, Lecturer, 衛生化学研究室, 講師 (30297171)
FUJISHIMA Toshie Teikyo University Faculty of Pharmaceutical Sciences, Lecturer, 薬学部, 講師 (90286980)
SUGIYAMA Toru The University of Tokyo Department of Life Sciences, Assistant Professor, 大学院・総合文化研究科, 助手 (40242036)
吉田 彰宏 帝京大学, 薬学部, 助手 (10292301)
|
|---|
| Project Period (FY) |
2001 – 2002
|
| Project Status |
Completed (Fiscal Year 2002)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | RecA Protein / gene recombination enzyme / oligonucleotides / Schiff base / NFκB / nuclear receptor / active vitamin D3 / super agonist / CDスペクトル / ゲル電気泳動 / 凝集 / ホルミルウリジン |
|---|
| Research Abstract |
5-Formy-2'-deoxyluidine (fdU) is an oxidative thymidine lesion, which has been known to be mutagenic and cytotoxic in vivo. Whereas these unfavolable biological effects of fU appear to be largely caused by mispairing during replication and transcription and inhibition of metabolic enzymes, it is intriguing to consider the reaction of formyl group of fU with proteins affording cross-link and its implications for the biological effects. We showed that single-stranded oligonucleotides containing 5-formyl-2'-deoxyuridine (dfU) can cross-link the peptides derived from the DNA binding site of RecA protein, extending from residues 193 to 212, through a Schiff base formation. The ability of cross-linking of fdU-containing oligonucleotides was investigated using a series of peptides whose amino acid residues spanning the center of the RecA-derived peptide were sequentially replaced with lysine. Circular dichroism (CD) spectroscopy and gel mobility shift assay demonstrated that cross-linking reaction proceeded effici ntly only when the peptides bound to the oligonucleotides. Furthermore, sedimentation experiment revealed that the final cross-linked products were in state of aggregation. These results demonstrate that fdU-containing oligonucleotides would be a useful tool for identifying lysines in close proximity to the nucleobase in protein-DNA complexes. RecA protein could be a useful carrier in transporting modified nucleotides to the target gene. We also investigated the other transcription factors of NFκB and vitamin D receptor.
|
