Trinh-Rohlik Q, Maxwell E S
Department of Biochemistry, North Carolina State University, Raleigh 27695-7622.
Nucleic Acids Res. 1988 Jul 11;16(13):6041-56. doi: 10.1093/nar/16.13.6041.
Previous work has reported the isolation and sequencing of a mouse low molecular weight RNA species designated 4.5S hybridizing RNA or hybRNA because of its ability to intermolecularly hybridize with mouse mRNA and 18S rRNA sequences. Using synthetic DNA oligonucleotide probes we have examined the conservation of this gene sequence and its expression as a lmwRNA transcript across evolution. Southern blot analysis has shown that homologous genes of single or low copy number are found in all eukaryotes examined as well as in E. coli. Northern blot analysis has demonstrated 4.5S hybRNA transcription in all mouse tissues as well as expression in yeast and Xenopus laevis as lmwRNAs of approximately 130 and 100 nucleotides, respectively, as compared with mouse/rat/hamster species of approximately 87 nucleotides. Yeast and X. laevis 4.5S hybRNA homologs, isolated by hybrid-selection, were shown by Northern blot analysis to intermolecularly hybridize with homologous as well as heterologous 18S rRNA sequences. The conservation of 4.5S hybRNA homologous genes and their expression as lmwRNA transcripts with common intermolecular RNA:RNA hybridization capabilities in fungi, amphibians, and mammals argues for a common, conserved and required biological function for this lmwRNA in all eukaryotes and potential utilization of its intermolecular RNA:RNA hybridization capabilities to carry out this function.
先前的研究报道了一种小鼠低分子量RNA的分离和测序,该RNA被命名为4.5S杂交RNA或hybRNA,因其能够与小鼠mRNA和18S rRNA序列进行分子间杂交。我们使用合成的DNA寡核苷酸探针,研究了该基因序列在进化过程中的保守性及其作为低分子量RNA转录本的表达情况。Southern印迹分析表明,在所检测的所有真核生物以及大肠杆菌中都发现了单拷贝或低拷贝数的同源基因。Northern印迹分析显示,4.5S hybRNA在所有小鼠组织中都有转录,在酵母和非洲爪蟾中也有表达,分别为大约130和100个核苷酸的低分子量RNA,而小鼠/大鼠/仓鼠的该RNA约为87个核苷酸。通过杂交筛选分离得到的酵母和非洲爪蟾4.5S hybRNA同源物,经Northern印迹分析表明,它们能够与同源以及异源的18S rRNA序列进行分子间杂交。4.5S hybRNA同源基因的保守性及其作为具有共同分子间RNA:RNA杂交能力的低分子量RNA转录本的表达,表明这种低分子量RNA在所有真核生物中具有共同的、保守的和必需的生物学功能,并且可能利用其分子间RNA:RNA杂交能力来执行这一功能。