Shared protein components of SINE RNPs.
Shared protein components of SINE RNPs.
复制标题
SINE RNP 的共享蛋白质成分。
DOI：
10.1016/s0022-2836(02)00542-9
复制
发表时间：
2002
影响因子：
5.6
通讯作者：
Deininger,Prescott
中科院分区：
文献类型：
--
作者：
West,Neva;Roy-Engel,AstridM;Imataka,Hiroaki;Sonenberg,Nahum;Deininger,Prescott
由高拷贝短移动的元件（西内斯）产生的异质短RNA与蛋白质相互作用形成RNA-蛋白质（RNP）复合物。特别是，BC 1 RNA，其在脑和睾丸中特异性地从ID SINE家族的一个基因座转录到高水平，作为离散的RNP复合物存在。我们在几种细胞系中表达了一系列改变的BC 1和其他SINE相关RNA，并在非变性PAGE测定中测试了所得RNP复合物的迁移率，以确定这些SINE RNA的哪些部分有助于蛋白质结合。当不同的SINE RNA取代BC 1 ID序列时，所得RNP表现出与BC 1相同的迁移率。这表明与BC 1的ID部分结合的蛋白质不是序列特异性的，并且可能更依赖于RNA的二级结构。这也表明所有的SINE RNA都可以结合一组相似的细胞蛋白。缺失BC 1 RNA的富含A的区域对RNP的移动性具有显著影响。啮齿动物细胞系表现出轻微不同的流动性，这种转变复杂的人细胞系相比，反映了一个或多个蛋白质组分的进化差异。基于在RNP复合物中观察到的当去除富含A的区域时的迁移率变化，我们决定检查多聚（A）结合蛋白（PABP）作为RNP的候选成员。针对PABP的C末端的抗体能够免疫沉淀BC 1 RNA，证实PABP存在于BC 1 RNP中。鉴于poly（A）区域在反转录转座过程中的普遍作用，这些数据表明PABP可能有助于SINE反转录转座过程。
The heterogeneous, short RNAs produced from the high, copy, short mobile elements (SINEs) interact with proteins to form RNA–protein (RNP) complexes. In particular, the BC1 RNA, which is transcribed to high levels specifically in brain and testis from one locus of the ID SINE family, exists as a discrete RNP complex. We expressed a series of altered BC1, and other SINE-related RNAs, in several cell lines and tested for the mobility of the resulting RNP complexes in a native PAGE assay to determine which portions of these SINE RNAs contribute to protein binding. When different SINE RNAs were substituted for the BC1 ID sequence, the resulting RNPs exhibited the same mobility as BC1. This indicates that the protein(s) binding to the ID portion of BC1 is not sequence specific and may be more dependent upon the secondary structure of the RNA. It also suggests that all SINE RNAs may bind a similar set of cellular proteins. Deletion of the A-rich region of BC1 RNA has a marked effect on the mobility of the RNP. Rodent cell lines exhibit a slightly different mobility for this shifted complex when compared to human cell lines, reflecting evolutionary differences in one or more of the protein components. On the basis of mobility change observed in RNP complexes when the A-rich region is removed, we decided to examine poly(A) binding protein (PABP) as a candidate member of the RNP. An antibody against the C terminus of PABP is able to immunoprecipitate BC1 RNA, confirming PABP's presence in the BC1 RNP. Given the ubiquitous role of poly(A) regions in the retrotransposition process, these data suggest that PABP may contribute to the SINE retrotransposition process.