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

我们已经通过使用如下分别描述的两种研究系统来进行我们的研究项目：（1）使用NRK（正常大鼠肾）细胞：NRK细胞是有用的，因为它们通过加入EGF和TGF-β而可逆地转化。我们已经建立了几种突变株，它们对EGF和TGF-β的作用不敏感。利用这些突变体，我们研究了EGF和几个癌基因的信号通路和锚定依赖性。(2)利用粟酒裂殖酵母细胞分裂周期（cdc）突变体cdc 2、cdc 13和cdc 10：我们克隆并分析了3个新的人类基因，它们与cdc 2和cdc 13互补。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cdc 13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测序结果表明，这两个克隆编码的蛋白具有两个锚蛋白基序，且与cdc 10基因产物和SWI 4/SW 16（酿酒酵母）基因产物具有同源结构域。发现一个克隆是cdc 18本身。一个克隆（rep 1）编码一个锌指蛋白，这是需要在减数分裂前DNA合成。一个克隆（HAC 1）是ATF/CREB的同源物。其余三个克隆的DNA序列有待确定。

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.