Kastner B, Lührmann R
Max-Planck-Institut für Molekulare Genetik, Otto Warburg Laboratories, Berlin, FRG.
EMBO J. 1989 Jan;8(1):277-86. doi: 10.1002/j.1460-2075.1989.tb03374.x.
Small nuclear ribonucleoprotein particles U1 (U1 snRNPs) from HeLa cells were purified to homogeneity and examined in the electron microscope by negative staining with the double carbon film technique. A survey of several hundred views of the U1 snRNP particles revealed a main body about 8 nm in diameter with two characteristic protuberances 4-7 nm long and 3-4 nm wide. Anti-m3G antibodies were then used to define the site of the m3G base in the 5' cap of the U1 snRNA, close to the site where base-pairing occurs with the 5' splice sequence of pre-mRNA. The m3G base is located on the body of the U1 snRNP, close to one of the protuberances and approximately opposite the other.
从HeLa细胞中纯化出小核核糖核蛋白颗粒U1(U1 snRNPs),使其达到均一状态,并采用双碳膜技术进行负染,在电子显微镜下观察。对数百个U1 snRNP颗粒视图的调查显示,其主体直径约为8纳米,有两个特征性突起,长4 - 7纳米,宽3 - 4纳米。然后使用抗m3G抗体来确定U1 snRNA 5'帽中m3G碱基的位置,该位置靠近与前体mRNA的5'剪接序列发生碱基配对的位点。m3G碱基位于U1 snRNP的主体上,靠近其中一个突起,且大致与另一个突起相对。