Buvoli M, Cobianchi F, Bestagno M G, Mangiarotti A, Bassi M T, Biamonti G, Riva S
Istituto di Genetica Biochimica ed Evoluzionistica CNR, Pavia, Italy.
EMBO J. 1990 Apr;9(4):1229-35. doi: 10.1002/j.1460-2075.1990.tb08230.x.
The human hnRNP core protein A1 (34 kd) is encoded by a 4.6 kb gene split into 10 exons. Here we show that the A1 gene can be differentially spliced by the addition of an extra exon. The new transcript encodes a minor protein of the hnRNP complex, here defined A1B protein, with a calculated mol. wt of 38 kd, that coincides with a protein previously designated as B2 by some authors. In vitro translation of the mRNAs selected by hybridization with A1 cDNA produced two proteins of 34 and 38 kd; Northern blot analysis of poly(A)+ RNA from HeLa cells revealed that the abundance of the A1B mRNA was approximately 5% that of A1. The A1B protein was detected by Western blotting with an anti-A1 monoclonal antibody both in enriched preparations of basic hnRNP proteins and in 40S hnRNP particles. The A1B protein exhibits a significantly higher affinity than A1 for ssDNA. The recombinant A1B protein, expressed in Escherichia coli, shows the same electrophoretic mobility and charge as the cellular one.
人类不均一核糖核蛋白核心蛋白A1(34千道尔顿)由一个4.6千碱基对的基因编码,该基因被分成10个外显子。在此我们表明,通过添加一个额外的外显子,A1基因可以进行差异剪接。新的转录本编码一种不均一核糖核蛋白复合物的次要蛋白,在此定义为A1B蛋白,计算分子量为38千道尔顿,它与一些作者先前指定为B2的一种蛋白一致。用A1 cDNA杂交选择的mRNA进行体外翻译产生了34千道尔顿和38千道尔顿的两种蛋白;对来自HeLa细胞的聚腺苷酸加尾RNA进行Northern印迹分析表明,A1B mRNA的丰度约为A1的5%。用抗A1单克隆抗体通过蛋白质免疫印迹法在碱性不均一核糖核蛋白的富集制剂和40S不均一核糖核蛋白颗粒中均检测到了A1B蛋白。A1B蛋白对单链DNA的亲和力比A1显著更高。在大肠杆菌中表达的重组A1B蛋白与细胞内的A1B蛋白表现出相同的电泳迁移率和电荷。