Molecular cloning of cDNA for the nuclear ribonucleoprotein particle C proteins: a conserved gene family.
Molecular cloning of cDNA for the nuclear ribonucleoprotein particle C proteins: a conserved gene family.
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核核糖核蛋白颗粒 C 蛋白 cDNA 的分子克隆：保守基因家族。
DOI：
10.1073/pnas.83.7.2007
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发表时间：
1986
影响因子：
11.1
通讯作者：
Dreyfuss,G
中科院分区：
文献类型：
--
作者：
Nakagawa,TY;Swanson,MS;Wold,BJ;Dreyfuss,G
C蛋白C1和C2是脊椎动物异质核RNA （hnRNA）核糖核蛋白（hnRNP）复合物的主要成分。C1和C2是抗原相关的磷酸化蛋白，在完整细胞中与hnRNA接触，并在体外与RNA紧密结合。免疫筛选含单克隆抗体的人lambda gt11表达载体cDNA文库，分离出C蛋白cDNA克隆。lacZ-cDNA融合蛋白被两种不同的抗c蛋白单克隆抗体识别。与1.1千碱基长的cDNA克隆杂交选择的HeLa细胞mRNA在体外翻译得到C1和C2蛋白（分别为41和43 kDa）。RNA印迹分析显示，在人类细胞中，两个约1.4 kb和1.9 kb的多聚腺苷化转录本存在强杂交。基因组DNA印迹分析显示，人类和小鼠中存在多个杂交限制性内切片段，从人类到酵母的真核生物中均存在同源DNA序列。这些发现表明编码hnRNP C蛋白的序列是一个保守基因家族的成员，它们为这些蛋白的详细分子和遗传学研究开辟了道路。
The C proteins, C1 and C2, are major constituents of the heterogeneous nuclear RNA (hnRNA) ribonucleoprotein (hnRNP) complex in vertebrates. C1 and C2 are antigenically related phosphoproteins that are in contact with hnRNA in intact cells and bind to RNA tightly in vitro. A cDNA clone for the C proteins was isolated by immunological screening of a human lambda gt11 expression vector cDNA library with monoclonal antibodies. The lacZ-cDNA fusion protein is recognized by two different anti-C protein monoclonal antibodies. HeLa cell mRNA that was hybrid-selected with the cDNA clone (1.1 kilobases long) was translated in vitro and yielded both the C1 and C2 proteins (41 and 43 kDa, respectively). RNA blot analysis showed strong hybridization to two polyadenylylated transcripts, of about 1.4 kb and 1.9 kb, in human cells. Genomic DNA blot analysis showed multiple hybridizing restriction fragments in human and mouse, and homologous DNA sequences are found across eukaryotes from human to yeast. These findings suggest that the sequences encoding the hnRNP C proteins are members of a conserved gene family and they open the way for detailed molecular and genetic studies of these proteins.