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.

我们通过使用两种研究系统进行了研究，如下所述；（1）使用NRK（正常大鼠肾脏）细胞：NRK细胞很有用，因为它们通过添加EGF和TGF-beta会可逆地转化。我们已经建立了几种对EGF和TGF-beta效果不敏感的突变菌株。使用这些突变体，我们研究了EGF和几种肿瘤基因的信号途径以及锚定依赖性。（2）使用S.Pombe细胞分裂周期（CDC）突变体，CDC2，CDC13和CDC10：我们已经克隆并分析了3个补充CDC2和CDC13的3个新型人类基因。 DNA测序表明它们编码RNA结合蛋白。他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。我们还克隆了补充CDC 10突变体的8种新型基因。 DNA测序揭示了两个克隆用两个Ankyrin基序编码蛋白质，并具有与Cdc10基因产物以及SWI4/SW16（S.cerevisiae）基因产物的同源域的蛋白质。发现一个克隆本身是CDC18。一个克隆（REP1）编码在前DNA合成中需要的锌指蛋白。一个克隆（HAC1）是ATF/CREB的同源物。三个克隆其余的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、

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。”

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。”

Tamura K, Kanaoka Y, Nojima H, Okayama H: Cyclin G: "a new cyclin with a potential tyrosine phosphorylation site." Oncogene.

Tamura K、Kanaoka Y、Nojima H、Okayama H：细胞周期蛋白 G：“一种具有潜在酪氨酸磷酸化位点的新细胞周期蛋白。”