Project/Area Number |
01580199
|
Research Category |
Grant-in-Aid for General Scientific Research (C)
|
Allocation Type | Single-year Grants |
Research Field |
代謝生物化学
|
Research Institution | Keio university School of Medicine |
Principal Investigator |
TAKANO Toshiya Keio University School of Medicine, Department of Microbiology, Professor, 医学部, 教授 (60051364)
|
Co-Investigator(Kenkyū-buntansha) |
TACHIBANA Kouichi Keio University School of Medicine, Department of Microbiology, Instructor, 医学部, 助手 (80216986)
瀬川 薫 慶應義塾大学, 医学部, 講師 (30114523)
|
Project Period (FY) |
1989 – 1991
|
Project Status |
Completed (Fiscal Year 1991)
|
Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
|
Keywords | Rab-2 gene / GTP-binding protein / Transcription / Complementary DNA / Messenger RNA / Polyadenylation site / Antiserum / Modification of protein / GTP結合蛋白質 / cDNA / 蛋白質のりん酸化 / rabー2蛋白質 / ウエスタンブロック法 / 免疫沈降法 / 蛍光抗体法 / 組織特異的発現 / poly A 付加部位 / houseーkeeping gene / 第8染色体 / 腎糸球体 / 腎尿細管 / 転写 / house-keeping gene / 脳 / ウエスタンブロット分析 / 3'非翻訳領域 |
Research Abstract |
The human rab-2 gene codes a 24-kDa YPT1-related GTP-binding protein. In all the human cell lines examined, including hematopoietic, fibroblastic and tumor cells, three rab-2 transcripts of 3.5, 2.4 and 1.4 kb were detected. The 2.4- and 3.5-kb rab-2 RNA hybridized with the probe specific for the 3'-non-coding region of 2.4-kb transcript, but not the 1.4-kb RNA. These transcripts seemed to differ in the length of their 3'-non-translated regions due to terminations of the transcription and/or processing of the primary transcript at the alternative polyadenylation sites. The smaller 1.4-kb transcript was the most predominant in Molt-4 and U-937, while the 2.4-kb RNA was dominant in the other cell lines. The degradation rates of all the rab-2 transcripts wbre similarly as slow as beta-actin RNA, even in the two cell lines whose predominant species of the transcripts were different. Rab-2 protein was detected in the extracts of human cultured cells as one major band of 24 kDa and a more faint band of 25 kDa by both immunoblotting and immunoprecipitation. Only the 25-kDa protein was detected in HeLa and KOBK101 cells. In the COS-1 cells transfected with an expression vector carrying rab-2 cDNA, the rab-2 protein was mainly located in the cytoplasm.
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