Ohlsson R I, van Eekelen C, Philipson L
Nucleic Acids Res. 1982 May 25;10(10):3053-68. doi: 10.1093/nar/10.10.3053.
Heterogeneous nuclear protein complexes (hnRNP) containing the precursor RNA from the adenovirus early region 2 were analysed to determine the specificity of protein-RNA interaction. RNA precursor sequences were present in isolated hnRNP complexes and endogenous 30S particles. At least 20-40 bases long fragments were protected when RNase A was used to remove unprotected RNA sequences in hnRNA complexes. Similarly around 40 bases of RNA were protected in 30S particles. These sequences represent discrete regions of the adenovirus genome. Especially sequences complementary to the EcoRI-F fragment encoding the first leader and the major intron for the DNA binding protein (DBP) RNA precursor, were analysed in detail. Tentatively, sequences resistant to RNase A were located in the middle of the intron and at the splice-donor junction of the first leader of the DBP precursor RNA. The same sequences were identified irrespective whether hnRNP complexes or 30S particles were used suggesting that 30S particles originate from hnRNP complexes. A 38.000 dalton protein appears to be in direct contact with RNA sequences complementary to the EcoRI-F fragment.
对含有腺病毒早期区域2前体RNA的异质核蛋白复合物(hnRNP)进行分析,以确定蛋白质-RNA相互作用的特异性。RNA前体序列存在于分离的hnRNP复合物和内源性30S颗粒中。当使用核糖核酸酶A去除hnRNA复合物中未受保护的RNA序列时,至少20 - 40个碱基长的片段受到保护。同样,在30S颗粒中约40个碱基的RNA受到保护。这些序列代表腺病毒基因组的离散区域。特别是对与编码第一个前导序列和DNA结合蛋白(DBP)RNA前体的主要内含子的EcoRI - F片段互补的序列进行了详细分析。初步确定,对核糖核酸酶A有抗性的序列位于内含子中间以及DBP前体RNA第一个前导序列的剪接供体连接处。无论使用hnRNP复合物还是30S颗粒,都鉴定出相同的序列,这表明30S颗粒起源于hnRNP复合物。一种38000道尔顿的蛋白质似乎与与EcoRI - F片段互补的RNA序列直接接触。