Yuan Y, Reddy R
Baylor College of Medicine, Department of Pharmacology, Houston, TX 77030.
Biochim Biophys Acta. 1989 Jun 1;1008(1):14-22. doi: 10.1016/0167-4781(89)90164-4.
Six human genomic clones containing sequences homologous to the U3 small nuclear RNA (snRNA) were isolated and characterized. Four of these clones were real U3 snRNA genes because they were transcribed in frog oocytes and the DNA sequences corresponding to the U3 snRNA were identical to the U3 snRNA of HeLa cells. The nucleotide sequences of four true U3 snRNA genes, 537 nucleotides on the 5'-flanking region and 340 nucleotides on the 3'-flanking region, were found to be identical. In addition, the restriction patterns, upto 2 kb on the 5' side and 2.2 kb on the 3' side, appeared to be same. All the isolated U3 clones, containing 15-20 kb of genomic DNA, contained only one U3 snRNA gene, indicating that the human U3 snRNA genes are several kilobases apart. One of the U3 clones contained a full-length U3 pseudogene. Southern blot analysis of genomic DNA with cloned U3 DNA as probe indicated that human DNA contains two families of U3 genes which differ in their flanking sequences. In the 5' flanking region of human U3 snRNA genes, homology to U-gene promoter element, an octamer motif, the 'U3 box', SP1 binding sites and a consensus 3' box in the 3' flanking region, were observed. These data show that the genomic organization and the sequence motifs that control transcription of human nucleolar U3 snRNA genes are similar to those of human U1 and U2 snRNA genes and suggest common mechanism(s) in the evolution of snRNA genes.
分离并鉴定了六个含有与U3小核RNA(snRNA)同源序列的人类基因组克隆。其中四个克隆是真正的U3 snRNA基因,因为它们在蛙卵母细胞中被转录,并且与U3 snRNA对应的DNA序列与HeLa细胞的U3 snRNA相同。发现四个真正的U3 snRNA基因的核苷酸序列,5'侧翼区域的537个核苷酸和3'侧翼区域的340个核苷酸,是相同的。此外,5'侧长达2 kb和3'侧长达2.2 kb的限制性酶切图谱似乎也是相同的。所有分离的U3克隆,包含15 - 20 kb的基因组DNA,只含有一个U3 snRNA基因,这表明人类U3 snRNA基因相隔数千个碱基对。其中一个U3克隆包含一个全长U3假基因。以克隆的U3 DNA为探针进行基因组DNA的Southern印迹分析表明,人类DNA包含两个侧翼序列不同的U3基因家族。在人类U3 snRNA基因的5'侧翼区域,观察到与U基因启动子元件、八聚体基序、“U3框”、SP1结合位点以及3'侧翼区域的共有3'框的同源性。这些数据表明,人类核仁U3 snRNA基因的基因组组织和控制转录的序列基序与人类U1和U2 snRNA基因相似,并提示了snRNA基因进化中的共同机制。