| Project/Area Number |
11670988
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Hematology
|
|---|
| Research Institution | Fukui Medical University |
|---|
Principal Investigator |
YOSHIDA Akira Fukui Medical University, Dept. of Medicine, Assistant Professor, 医学部, 助手 (80252005)
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
|
|---|
| Keywords | Apoptosis / endonuclease / AN34 / AP-ENDONUCLEASE / Caspase-3 / 白血病細胞 / エンドヌクレアーゼ / Caspase 3 / AN-34 / Caspase3 |
|---|
| Research Abstract |
We previously purified an apoptotic nuclease 34kDa (AN34) from human leukemia HL-60 cells (Cancer Res 58,1998). AN34 is a Mg2+ dependent endonuclease which induces DNA single and double strand breaks. In this study, we investigated whether AN34 is regulated by caspase 3 using cell-free system and chromatography. AN34 was purified from HL-60 cell lysate which was preincubated with recombinant caspase 3. DFF40/CAD was also separated from caspase 3 treated cell lysate at different fraction of ion exchange column. However, we could not isolate AN34 and DFF40/CAD from non-treated cell lysates. We compared biochemical property of AN34 with DFF40/CAD. AN34 predominantly induces DNA single strand breaks. On the other hand, DFF40/CAD induces mainly DNA doubles strand breaks. In addition, AN34 showed not only endonuclease activity but also strong 3'-5'exonuclease activity. These data indicate that AN34 is a unique caspase 3 regulated 3'-5'exo-endonuclease which shows distinct biochemical property from DFF40/CAD.
|
