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Comprehensive isolation and functional analysis of the genes whose expressions are specifically induced upon entry into the Go phase of cell cycle

对进入细胞周期 Go 期时特异性诱导表达的基因进行全面分离和功能分析

基本信息

批准号：
12794010
负责人：
NOJIMA Hiroshi
金额：
$ 22.53万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for University and Society Collaboration
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

Mammalian cells stops growth and enter into the static phase or G_0 phase when the concentration of fetal calf serum in the culture medium was reduced from 10% to 0.5%. It was believed that a certain proliferation inhibitory factors exist in the medium harboring the cells in the G_0 phase to maintain the static condition. However, very few studies have been reported on the genes and their gene products that define the G_0 phase. Thus, if a large scale isolation of the genes whose expression was induced is performed, it would be possible to isolate the gene that is required to maintain the static phase by growth-inhibitory function. To isolate such genes comprehensively, we used an improved cDNA library subtraction p as we described previously. Using this method, we have so far been able to isolate more than 60 genes whose expression is induced in growth arrest condition and termed the transcript induced in growth arrest, TIGA, genes. Transcription of some of the TIGA genes are also induced upon contact inhibition of the cell growth. One of the TIGA genes, TIGA1, encodes a small protein with a single transmembrane motif. Ecopically expressed Tiga1 potently inhibited cell growth. Functional analyses of these TIGA genes might shed light to understanding the molecular mechanisms of induction and maintenance of G_0 state of the cell cycle.

当培养基中胎牛血清浓度从10%降至0.5%时，哺乳动物细胞停止生长，进入静止期或G_0期。人们认为，G_0期细胞的培养基中存在一定的增殖抑制因子，以维持静止状态。然而，关于定义G_0期的基因及其基因产物的研究报道很少。因此，如果对表达被诱导的基因进行大规模分离，则可以分离通过生长抑制功能维持静止期所需的基因。为了全面分离这些基因，我们使用了改进的 cDNA 文库扣除 p，如前所述。使用这种方法，我们迄今为止已经能够分离出60多个在生长停滞条件下诱导表达的基因，并将其称为生长停滞诱导的转录本，TIGA基因。接触抑制细胞生长时也会诱导一些 TIGA 基因的转录。 TIGA 基因之一 TIGA1 编码具有单个跨膜基序的小蛋白质。异位表达的 Tiga1 有效抑制细胞生长。这些 TIGA 基因的功能分析可能有助于理解细胞周期 G_0 状态诱导和维持的分子机制。