基因-蛋白质屏幕揭示了环素D1在发育中的转录作用
Frédéric Bienvenu1, Siwanon Jirawatnotai, Joshua E Elias
1Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.
Nature
|January 22, 2010
概括
细胞循环蛋白Cyclin D1在小鼠发育过程中也在转录中发挥作用. 它将CBP招募到Notch1基因促进体,调节其表达并影响视网膜前代细胞的增殖.
科学领域:
- 分子生物学分子生物学
- 发展生物学 发展生物学
- 癌症研究 癌症研究
背景情况:
- 环素D1是一种关键的细胞循环调节剂,在人类癌症中经常过度表达.
- 在正常发育和瘤发生过程中,cyclin D1的完整功能在很大程度上是未知的.
研究的目的:
- 为了研究在细胞循环中的作用之外的cyclin D1的体内功能.
- 通过基因-蛋白质基因方法识别新型环林D1结合蛋白和转录标.
主要方法:
- 开发Flag和血凝素标记的cyclin D1敲进小鼠模型.
- 高通量质谱检测用于识别环林D1相互作用蛋白.
- 染色体免疫沉与DNA微阵列 (ChIP-chip) 结合,用于全基因组位置分析.
主要成果:
- 发现Cyclin D1在小鼠发育过程中结合了活跃转录基因的促进者.
- 在发育中的视网膜中,cyclin D1将CREB结合蛋白 (CBP) 招募到Notch1基因促进体中.
- 失去D1环素导致CBP招募减少,Notch1表达减少,视网膜原生细胞增殖受损.
结论:
- 环素D1在小鼠发育中具有显著的体内转录功能,独立于其规范细胞周期的作用.
- 已确定的"遗传蛋白质学"方法是研究各种蛋白质体内功能的一种多功能工具.
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