Yurkova M S, Murray M T
Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, Michigan 48202, USA.
J Biol Chem. 1997 Apr 18;272(16):10870-6. doi: 10.1074/jbc.272.16.10870.
In oocytes, nontranslated maternal mRNAs are packaged by protein into messenger ribonucleoprotein particles (mRNPs) that are masked from translation by protein-RNA interactions. Proteins associated with such masked states of mRNAs are particularly abundant in amphibian oocytes. One of these mRNP proteins from Xenopus oocytes, mRNP3+4 (also called FRG Y2a/b or p54/p56), binds to diverse mRNAs independent of their sequence and is the germ line member of the evolutionarily conserved Y box protein multigene family. Xenopus oocytes contain soluble pools of mRNP3+4 6 S oligomers, probably dimers, and larger approximately 15 S particles containing mRNP3+4 and additional proteins. Here we report the purification of this larger form as an approximately 320-kDa particle that contains mRNP3+4 and nine additional polypeptides, including mRNA-binding polypeptides of 34 and 36 kDa and a doublet of 110/105 kDa that proved to be nucleolin. The particle has a protein kinase activity that phosphorylates its own mRNP3+4, nucleolin, and a 31-kDa polypeptide component and exhibits translational inhibition in both the wheat germ extract and rabbit reticulocyte lysate systems. The presence of mRNP3+4 and nucleolin in this large translation regulatory particle suggests that it participates in an early step of mRNP assembly and masking.
在卵母细胞中,未翻译的母体mRNA由蛋白质包装成信使核糖核蛋白颗粒(mRNP),这些颗粒因蛋白质-RNA相互作用而被阻止翻译。与这种mRNA的掩盖状态相关的蛋白质在两栖类卵母细胞中特别丰富。非洲爪蟾卵母细胞中的一种mRNP蛋白,即mRNP3 + 4(也称为FRG Y2a / b或p54 / p56),能独立于mRNA序列与多种mRNA结合,并且是进化上保守的Y盒蛋白多基因家族的生殖系成员。非洲爪蟾卵母细胞含有mRNP3 + 4 6S寡聚体的可溶性池,可能是二聚体,以及含有mRNP3 + 4和其他蛋白质的大约15S的较大颗粒。在这里,我们报告将这种较大形式纯化成为一个约320 kDa的颗粒,它包含mRNP3 + 4和另外九种多肽,包括34 kDa和36 kDa的mRNA结合多肽以及经证实为核仁素的110/105 kDa的双联体。该颗粒具有蛋白激酶活性,可磷酸化其自身的mRNP3 + 4、核仁素和一种31 kDa的多肽成分,并且在小麦胚芽提取物和兔网织红细胞裂解物系统中均表现出翻译抑制作用。这种大型翻译调节颗粒中存在mRNP3 + 4和核仁素,表明它参与了mRNP组装和掩盖的早期步骤。