2005 Fiscal Year Final Research Report Summary
Chemical and Biochemical Studies on Pathological Aspect of Nitric Oxide
| Project/Area Number |
15350099
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Chemistry related to living body
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
MAKINO Keisuke Kyoto University, International Innovation Center, Professor, 国際融合創造センター, 教授 (50159141)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KODAKI Tsutomu Kyoto University, Institute ofAdvanced Energy, Associate Professor, エネルギー理工学研究所, 助教授 (70170264)
KANAORI Kenji Kyoto Institute of Technology, Graduate School of Science and Technology, Associate Professor, 大学院工芸科学研究科, 助教授 (30273543)
|
|---|
| Project Period (FY) |
2003 – 2005
|
| Keywords | Nitric Oxide / Carcinogenesis / Deoxvoxanosine (dOxo) / dOxo amidite monomer synthesis / Synthesis of DNA oligomer containing dOxo / Crosslink formation between dOxo in DNA and repair enzymes / Enzymes repairing dOxo in DNA / Recognition of dOxo in DNA by DNA-relevant enzymes |
|---|
| Research Abstract |
Endogenous NO, necessary for many physiological functions, is sometimes overproduced in inflammation and therefore carcinogenic, however no clear evidence has been reported. In this study, we obtained important data to answer this essential question.
1. Oxanine (Oxa), a damaged base generated from Gua by NO-or HNO2-induced nitrosative deamination, has been considered as a mutagen-potent lesion. In this study, for exploring more detailed properties of Oxa, large-scale preparation of Oxa-containing DNA oligomers (Oxa-ODN) was developed: 2'-deoxynucleoside of Oxa (deoxyoxanosine, dOxo) obtained from dGuo by HNO2-nitrosation was subjected to 5'-0-selective tritylation (yield of DMT-dOxo, 70%). Subsequently DMT-dOxo was converted by conventional phosphoramidation to DMT-dOxo-amidite (yield, 72.5%). The amidite was used for synthesizing Oxa-ODNs: The coupling yields for Oxa incorporation and the total synthesis of 25mer DNA were over 93 and 85 %, respectively.
2. Using DNA oligomers with or wi
… More
thout dOxo as templates and primers, DNA polymerase chain elongation was investigated and we found that in the opposite site to Oxa, T and C are incorporated almost equally, indicating that Oxa is mutagenic.
3. We performed screening for the base-excision repair (BER) system to find no enzymic activity for the excision of Oxa, and revealed that Oxa-polyamine adduct is repaired by the nucleotide excision and recombination repair system, indicative of the cellular Oxa formation and that other DNA-relevant enzymes such as ligase and restriction enzymes recognize Oxa as Gua.
4. We also found that Oxa-recognizing BER enzymes cross-link with Oxa in the DNA duplex irreversibly and this activity is eliminated by heat, while histones binding DNA nonspecifically also crosslink Oxa in DNA duplexes although slowly. This unique crosslinking will be helpful for fishing Oxa-recognizing and-repairing enzymes.
5. NMR analysis for the structure around Oxa in DNA is now being conducted to reveal the mechanism for such unique properties of Oxa. Less
|
Research Products
(52 results)
-
-
-
-
-
-
-
-
-
[Journal Article] A hydrogen peroxide-generating agent, 6-formylpterin, enchances heat-induced apoptosis.2005
Author(s)
S.Wada, Z.-G.Cui, T.Kondo, Q.-L.Zhao, R.Ogawa, M.Shoji, T.Arai, K.Makino, I.Furuta
-
Journal Title
Int. J. Hyprethermia 21
Pages: 231-246
Description
「研究成果報告書概要(和文)」より
-
-
-
-
-
-
-
-
-
[Journal Article] A hydrogen peroxide-generating agent, 6-formylpterin, enhances heat-induced apoptosis.2005
Author(s)
S.Wada, Z.-G.Cui, T.Kondo, Q.-L.Zhao, R.Ogawa, M.Shoji, T.Arai, K.Makino, I.Furuta
-
Journal Title
Int. J. Hyprethermia 21
Pages: 231-246
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
