Nicoloso M, Caizergues-Ferrer M, Michot B, Azum M C, Bachellerie J P
Laboratoire de Biologie Moléculaire Eucaryote du CNRS, Université Paul-Sabatier, Toulouse, France.
Mol Cell Biol. 1994 Sep;14(9):5766-76. doi: 10.1128/mcb.14.9.5766-5776.1994.
We have found that intron 11 of the nucleolin gene in humans and rodents encodes a previously unidentified small nucleolar RNA, termed U20. The single-copy U20 sequence is located on the same DNA strand as the nucleolin mRNA. U20 RNA, which does not possess a trimethyl cap, appears to result from intronic RNA processing and not from transcription of an independent gene. In mammals, U20 RNA is an 80-nucleotide-long, metabolically stable species, present at about 7 x 10(3) molecules per exponentially growing HeLa cell. It has a nucleolar localization, as indicated by fluorescence microscopy following in situ hybridization with digoxigenin-labeled oligonucleotides. U20 RNA contains the box C and box D sequence motifs, hallmarks of most small nucleolar RNAs reported to date, and is immunoprecipitated by antifibrillarin antibodies. It also exhibits a 5'-3' terminal stem bracketing the box C-box D motifs like U14, U15, U16, or Y RNA. A U20 homolog of similar size has been detected in all vertebrate classes by Northern (RNA) hybridization with mammalian oligonucleotide probes. U20 RNA contains an extended region (21 nucleotides) of perfect complementarity with a phylogenetically conserved sequence in 18S rRNA. This complementarity is strongly preserved among distant vertebrates, suggesting that U20 RNA may be involved in the formation of the small ribosomal subunit like nucleolin, the product of its host gene.
我们发现,人类和啮齿动物的核仁素基因内含子11编码一种以前未被鉴定的小核仁RNA,称为U20。单拷贝的U20序列与核仁素mRNA位于同一条DNA链上。U20 RNA没有三甲基帽,似乎是由内含子RNA加工产生的,而不是来自独立基因的转录。在哺乳动物中,U20 RNA是一种长度为80个核苷酸的、代谢稳定的分子,在指数生长的HeLa细胞中,每个细胞约有7×10³个分子。原位杂交后用荧光显微镜观察发现,它定位于核仁。U20 RNA含有框C和框D序列基序,这是迄今为止报道的大多数小核仁RNA的特征,并且能被抗原纤维蛋白抗体免疫沉淀。它还像U14、U15、U16或Y RNA一样,在框C-框D基序两侧有一个5'-3'末端茎。通过与哺乳动物寡核苷酸探针进行Northern(RNA)杂交,在所有脊椎动物类别中都检测到了大小相似的U20同源物。U20 RNA包含一个与18S rRNA中系统发育保守序列完全互补的延伸区域(21个核苷酸)。这种互补性在远缘脊椎动物中得到了强烈保留,这表明U20 RNA可能像其宿主基因的产物核仁素一样,参与小核糖体亚基的形成。