Ogilvie V C, Wilson B J, Nicol S M, Morrice N A, Saunders L R, Barber G N, Fuller-Pace F V
Department of Molecular and Cellular Pathology, University of Dundee, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK.
Nucleic Acids Res. 2003 Mar 1;31(5):1470-80. doi: 10.1093/nar/gkg236.
The RNA helicases p68 and p72 are highly related members of the DEAD box family of proteins, sharing 90% identity across the conserved core, and have been shown to be involved in both transcription and mRNA processing. We previously showed that these proteins co-localise in the nucleus of interphase cells. In this study we show that p68 and p72 can interact with each other and self-associate in the yeast two-hybrid system. Co-immunoprecipitation experiments confirmed that p68 and p72 can interact in the cell and indicated that these proteins preferentially exist as hetero-dimers. In addition, we show that p68 can interact with NFAR-2, a protein that is also thought to function in mRNA processing. Moreover, gel filtration analysis suggests that p68 and p72 can exist in a variety of complexes in the cell (ranging from approximately 150 to approximately 400 kDa in size), with a subset of p68 molecules being in very large complexes (>2 MDa). The potential to exist in different complexes that may contain p68 and/or p72, together with a range of other factors, would provide the potential for these proteins to interact with different RNA substrates and would be consistent with recent reports implying a wide range of functions for p68/p72.
RNA解旋酶p68和p72是DEAD盒蛋白家族中高度相关的成员,在保守核心区域具有90%的同一性,并且已被证明参与转录和mRNA加工过程。我们之前表明这些蛋白在间期细胞的细胞核中共定位。在本研究中,我们表明p68和p72在酵母双杂交系统中可以相互作用并自我缔合。免疫共沉淀实验证实p68和p72在细胞中可以相互作用,并表明这些蛋白优先以异源二聚体形式存在。此外,我们表明p68可以与NFAR-2相互作用,NFAR-2是一种也被认为在mRNA加工中发挥作用的蛋白。而且,凝胶过滤分析表明p68和p72在细胞中可以存在于多种复合物中(大小范围从约150 kDa到约400 kDa),其中一部分p68分子存在于非常大的复合物中(>2 MDa)。存在于可能包含p68和/或p72以及一系列其他因子的不同复合物中的可能性,将为这些蛋白与不同RNA底物相互作用提供潜力,并且与最近暗示p68/p72具有广泛功能的报道一致。