Ficzycz Andrew, Ovsenek Nick
Department of Anatomy and Cell Biology, College of Medicine, 107 Wiggins Road, University of Saskatchewan, Saskatoon SK S7N 5E5, Canada.
J Biol Chem. 2002 Mar 8;277(10):8382-7. doi: 10.1074/jbc.M110304200. Epub 2001 Dec 4.
Yin Yang 1 (YY1) is a multifunctional transcription factor that activates, represses, or initiates transcription of a diverse assortment of genes. Previous studies suggest a role for YY1 in cellular growth and differentiation, but its biological function during development of the vertebrate oocyte or embryo remains to be determined. We recently showed that YY1 is abundantly expressed throughout oogenesis and early embryonic stages of Xenopus, but it is sequestered in the cytoplasm and does not function directly in transcriptional regulation. In the present study we used a series of biochemical analyses to explore the potential function of YY1 in the oocyte cytoplasm. YY1 was isolated from oocyte lysates by oligo(dT)-cellulose chromatography, suggesting that it associates with maternally expressed mRNA in vivo. RNA mobility shift assays demonstrate that endogenous YY1 binds to labeled histone mRNA. Size exclusion chromatography of oocyte lysates revealed that YY1 exists in high molecular mass complexes in the range of 480 kDa. Destruction of endogenous RNA by RNase treatment of lysates, abolished the binding of YY1 to oligo(dT)-cellulose and resulted in redistribution from 480-kDa complexes to the monomeric form. Microinjection of RNase directly into the cytoplasm released YY1 from 480-kDa complexes and unmasked its DNA-binding activity, but did not promote translocation to the nucleus. These results provide evidence that YY1 is a component of ribonucleoprotein (mRNP) complexes in the Xenopus oocyte, indicating a novel function for YY1 in the storage or metabolism of maternal transcripts.
阴阳1(YY1)是一种多功能转录因子,可激活、抑制或启动多种基因的转录。先前的研究表明YY1在细胞生长和分化中发挥作用,但其在脊椎动物卵母细胞或胚胎发育过程中的生物学功能仍有待确定。我们最近发现,YY1在非洲爪蟾的整个卵子发生和早期胚胎阶段都有大量表达,但它被隔离在细胞质中,并不直接参与转录调控。在本研究中,我们使用了一系列生化分析来探索YY1在卵母细胞细胞质中的潜在功能。通过寡聚(dT)-纤维素层析从卵母细胞裂解物中分离出YY1,这表明它在体内与母源表达的mRNA相关联。RNA迁移率变动分析表明,内源性YY1与标记的组蛋白mRNA结合。卵母细胞裂解物的尺寸排阻色谱显示,YY1以480 kDa范围内的高分子量复合物形式存在。用RNase处理裂解物破坏内源性RNA,消除了YY1与寡聚(dT)-纤维素的结合,并导致其从480 kDa复合物重新分布到单体形式。将RNase直接显微注射到细胞质中,使YY1从480 kDa复合物中释放出来,并揭示了其DNA结合活性,但并未促进其向细胞核的转运。这些结果提供了证据,表明YY1是非洲爪蟾卵母细胞核糖核蛋白(mRNP)复合物的一个组成部分,这表明YY1在母源转录本的储存或代谢中具有新功能。