| Project/Area Number |
05670300
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Immunology
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
HATTORI Masakazu Kyoto University, Faculty of Medicine, Dept.of Immunology and Cell Biology, Assistant Researcher, 医学部, 助手 (40211479)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MINATO Nagahiro Kyoto University, Faculty of Medicine, Dept.of Immunology and Cell Biology, Prof, 医学部, 教授 (40137716)
|
|---|
| Project Period (FY) |
1993 – 1994
|
| Project Status |
Completed (Fiscal Year 1994)
|
| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,700,000 (Direct Cost: ¥1,700,000)
|
|---|
| Keywords | RanGAP / RaplGAP / nuclear protein / cell cycle progression / Ran / RCC-1 system / Interleukin-2 / differntial hybridization / Ran GAP / 有糸分裂破綻 / リン酸化会合分子 / 細胞周期 / Rap1GAPホモログ / 翻訳時ホッピング / GAP活性 / 細胞増殖抑制 |
|---|
| Research Abstract |
We have cloned a novel cDNA (Spa-1) which was little expressed in the quiescent state but induced in the interleukin 2 (IL2) -stimulated cycling state of an IL2-responsive murine lymphoid cell line by differential hybridization. Spa-1 mRNA (3.5kb) was induced in the normal lymphocytes following various mitogenic stimulation. In normal organs it was preferentially expressed in both fetal and adult lymphohematopoietic tissues. A Spa-1-coded protein of 68 kDa was localized mostly in the nucleus. Its N-terminal domain is highly homologous to a human Rapl GTPase activating protein (GAP) , and a fusion protein of this domain (SpanN) indeed exhibited GAP activity for Rapl/Rsrl but not for Ras or Rho in vitro. Unlike the human Rapl GAP,however, SpanN also exhibited GAP activity for Ran, so far the only known Ras-related GTPase in the nucleus. In the presence of serum, stable Spa-1 cDNA transfectants of NIH3T3 (NIH/Spa-1) hardly overexpressed Spa-1 (p68) and grew normally as the parental cells. When NIH/Spa-1 cells were serum-starved to be arrested in G1/0, however, they exhibited progressive Spa-1 p68 accumulation unlike the control cells, and following the addition of serum they showed cell death resembling mitotic catastrophes of S phase during cell cycle progression. The results indicates that the novel nuclear protein, Spa-1, with a potentially active Ran GAP domain severely hampers the mitogen-induced cell cycle progression when abnormally and/or prematurely expressed. Functions of the Spa-1 protein and its regulation are discussed in the context of its possible interaction with Ran/RCC-1 system, which is involved in the coordinated nuclear functions including cell division.
|
