2012 Fiscal Year Final Research Report
Effects of DNA-protein cross-link lesions on eukaryotic transcription and replication
| Project/Area Number |
23710070
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Risk sciences of radiation/Chemicals
|
|---|
| Research Institution | Hiroshima University |
|---|
Principal Investigator |
NAKANO Toshiaki 広島大学, 大学院・理学研究科, 助教 (10526122)
|
|---|
| Project Period (FY) |
2011 – 2012
|
| Keywords | DNA損傷 / 転写 / 複製 |
|---|
| Research Abstract |
DNA-protein cross-links (DPCs) are superbulky and ubiquitous DNA lesions induced by various agents. It is likely that DPCs interfere with many aspects of DNA transactions. In the present study, we established in vitroassays to investigate the effect ofDPCs on eukaryotic transcription catalyzed by RNA polymerase II, showing that the formation of runoff products are dependent on the amount of nuclear cell extracts and reaction time. We then analyzed the effect of a model bulky lesion (fluorescein) on transcription. Fluorescein on the transcribed strand but not on the nontrascribed strand inhibited the transcription. We also established a novel method to introduce DPCs by employing Click reactions between protein azides and C5 alkyne-dT. We are currently studying the effect of DPCs on transcription by RNA polymerase II.
|
