Labhart P, Reeder R H
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Nucleic Acids Res. 1990 Sep 11;18(17):5271-7. doi: 10.1093/nar/18.17.5271.
In the ribosomal genes of X. laevis, the sequence GACTTGCNC is found about 60bp upstream of the gene promoter (T3) and is necessary and sufficient to cause termination of RNA polymerase I transcription. At the 3' end of the 40S precursor coding region (T2) a sequence differing by one nucleotide, GACTTGCNG, directs RNA 3' end formation but allows polymerase to transcribe on into the intergenic spacer (Labhart and Reeder, 1989, Genes and Dev. 4: 269-276). Sites corresponding to T2 and T3 are also found in a related species, X. borealis. Inspection of the T2 sequence in X. borealis reveals that it contains two copies of the terminator sequence, GACTTGCNC, located 15 and 96 bp downstream of the 3' end of the 40S precursor coding region. Here we present functional tests of those two T2 elements that show that, as predicted from the sequence, they both show termination activity and are functionally indistinguishable from the T3 site in X. laevis. These results suggest that X. laevis T2 is an example of a naturally occurring point mutation, and the inability to terminate transcription at T2 is an exception to the general pattern of ribosomal gene transcription in higher eukaryotes.
在非洲爪蟾的核糖体基因中,序列GACTTGCNC位于基因启动子(T3)上游约60bp处,是导致RNA聚合酶I转录终止所必需且充分的。在40S前体编码区(T2)的3'端,一个相差一个核苷酸的序列GACTTGCNG指导RNA 3'端的形成,但允许聚合酶继续转录进入基因间隔区(Labhart和Reeder,1989年,《基因与发育》4:269 - 276)。在相关物种北方爪蟾中也发现了与T2和T3对应的位点。对北方爪蟾T2序列的检查发现,它包含两个终止子序列GACTTGCNC的拷贝,位于40S前体编码区3'端下游15bp和96bp处。在此,我们展示了对这两个T2元件的功能测试,结果表明,正如从序列预测的那样,它们都表现出终止活性,并且在功能上与非洲爪蟾的T3位点没有区别。这些结果表明,非洲爪蟾的T2是自然发生的点突变的一个例子,而在T2处无法终止转录是高等真核生物核糖体基因转录一般模式的一个例外。