Cloning and analysis of the genes involved in the regulation of cell cycle and growth by complementation cloning.

通过互补克隆对参与细胞周期和生长调节的基因进行克隆和分析。

• 批准号: 02454537
• 负责人: NOJIMA Hiroshi
• 金额: $ 4.42万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454537/
• 关键词: Complementation; NRK; EGF; signal transduction; S.pombe; cdc mutant; Mlu motif; cAMP; NRK; EGF; シグナル伝達; pat1; 機能相補クロ-ニング; PDGF; transformation

We have pursued our research project by utilizing two kinds of research systems as described separately as follows;(1) Using NRK(normal rat kidney) cells: NRK cells are useful because they are transformed reversibly by the addition of EGF and TGF-beta. We have established several mutant strains which are insensitive to the effect of EGF and TGF-beta. Usingthese mutants, we have investigated the signal pathway of EGF and several oncogenes and anchorage dependency.(2) Using S.pombe cell division cycle (cdc) mutants, cdc2, cdc13 and cdc10: We have cloned and analysed 3 novel human genes that complement both cdc2 and cdc13. DNA sequencing revealed that they encode RNA binding proteins. They seem to complement these mutants by stabilizing and inducing translation of cdc13 gene product. We also cloned 8 kinds of novel genes that complement cdc 10 mutant. DNA sequencing unveiled that two clones encode proteins with two ankyrin motifs and with homologous domains to cdc10 gene product and those of SWI4/SW16 (S.cerevisiae) gene products. One clone was found to be cdc18 itself. One clone (rep1) encode a zinc-finger protein which is required in premeiotic DNA synthesis. One clone (HAC1) was a homologue of ATF/CREB. DNA sequence of the rest of three clones remain to be determined.

我们利用两种不同的研究系统进行我们的研究项目，分别如下：(1)使用正常大鼠肾脏细胞：NRK细胞是有用的，因为它们可以通过添加EGF和转化生长因子-β进行可逆转化。我们已经建立了几个对EGF和转化生长因子-β不敏感的突变株。利用这些突变体，我们研究了EGF和几个癌基因的信号通路以及锚定依赖性。(2)利用Pombe细胞分裂周期(CDC)突变体cdc2、cdc13和cdc10：我们克隆并分析了3个补充cdc2和cdc13的人类新基因。DNA测序显示，它们编码RNA结合蛋白。它们似乎通过稳定和诱导cdc13基因产物的翻译来补充这些突变体。我们还克隆了8种与CDC 10突变体互补的新基因。DNA测序表明，这两个克隆编码的蛋白具有两个锚定基序，并且与cdc10基因产物和SWI4/SW16(酿酒酵母)基因产物具有同源结构域。其中一个克隆被发现是cdc18本身。一个克隆(REP1)编码锌指蛋白，这是减数分裂前DNA合成所必需的。其中一个克隆(HAC1)与ATF/CREB同源。其余三个克隆的DNA序列仍有待确定。

项目成果

Tamura K,Kanaoka Y,Jinno S,Nagata A,Ogiso Y,Shimizu K,Hayakawa T,Nojima H, & Okayama H.: "Cyclin G: a new mammalian cyclin with a potential tyrosine phosphorylation site." Oncogene.

田村 K、金冈 Y、神野 S、永田 A、小木曾 Y、清水 K、早川 T、野岛 H、

Kizaka-Kondoh S,Sato K,tamura K,nojuma H, & Okayama H.: "Ralf-1 protein kinase is an integral component of oncogenic signal cascade shared by epidermal growth factor and platelet-derived growth factor," Molecular Cellular Biology. 12. 5078-5086 (1992)

木坂近藤 S、佐藤 K、田村 K、野朱马 H、

Tatsuka M, Mitsui H, Wada M, Nagata A, Nojima H, Okayama H: "Elongation factor-1alpha is a transformation-susceptibility determinant in BALB/c 3T3 cells." Nature. 359. 333-336 (1992)

Tatsuka M、Mitsui H、Wada M、Nagata A、Nojima H、Okayama H：“伸长因子 1α 是 BALB/c 3T3 细胞中的转化易感性决定因素。”

Tanaka K, Okazaki K, Okazaki N, Ueda T, Sugiyama A, Nojima H, Okayama H: "A new cdc gene required for S phase entry of Schizosaccharomyces pombe encodes a protein similar to the cdc10^+ and SWI4 gene products." EMBO J. 11(13). 4923-4932 (1992)

Tanaka K、Okazaki K、Okazaki N、Ueda T、Sugiyama A、Nojima H、Okayama H：“粟酒裂殖酵母进入 S 期所需的新 cdc 基因编码与 cdc10^ 和 SWI4 基因产物类似的蛋白质。”

Yagisawa H, Emori Y, Nojima H: "Phospholipase gene display restriction fragment length polymorphism between the genomes of normotensive and hypertensive rats." J Hypertens. 9(11). 303-307 (1991)

Yagisawa H、Emori Y、Nojima H：“磷脂酶基因在正常血压和高血压大鼠的基因组之间显示限制性片段长度多态性。”