Gudima S O, Taylor J M
Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, PA 19111-2497, USA.
BMC Biochem. 2001;2:3. doi: 10.1186/1471-2091-2-3. Epub 2001 Mar 21.
Previous studies by Volloch and coworkers have reported that during the expression of high levels of beta-globin mRNA in the spleen of anemic mice, they could also detect small but significant levels of an antisense (AS) globin RNA species, which they postulated might have somehow arisen by RNA-directed RNA synthesis. For two reasons we undertook to confirm and possibly extend these studies. First, previous studies in our lab have focussed on what is an unequivocal example of host RNA-directed RNA polymerase activity on the RNA genome of human hepatitis delta virus. Second, if AS globin species do exist they could in turn form double-stranded RNA species which might induce post-transcriptional gene silencing, a phenomenon somehow provoked in eukaryotic cells by AS RNA sequences.
We reexamined critical aspects of the previous globin studies. We used intraperitoneal injections of phenylhydrazine to induce anemia in mice, as demonstrated by the appearance and ultimate disappearance of splenomegaly. While a 30-fold increase in globin mRNA was detected in the spleen, the relative amount of putative AS RNA could be no more than 0.004%.
Contrary to earlier reports, induction of a major increase in globin transcripts in the mouse spleen was not associated with a detectable level of antisense RNA to globin mRNA.
沃洛克及其同事之前的研究报告称,在贫血小鼠脾脏中高水平β-珠蛋白mRNA表达期间,他们还能检测到少量但显著水平的反义(AS)珠蛋白RNA种类,他们推测这些可能是以某种方式通过RNA指导的RNA合成产生的。出于两个原因,我们着手确认并可能扩展这些研究。其一,我们实验室之前的研究集中在人类丁型肝炎病毒RNA基因组上宿主RNA指导的RNA聚合酶活性这一明确例子。其二,如果AS珠蛋白种类确实存在,它们可能反过来形成双链RNA种类,这可能会诱导转录后基因沉默,这是一种由AS RNA序列在真核细胞中以某种方式引发的现象。
我们重新审视了之前珠蛋白研究的关键方面。我们通过腹腔注射苯肼诱导小鼠贫血,脾肿大的出现和最终消失证明了这一点。虽然在脾脏中检测到珠蛋白mRNA增加了30倍,但假定的AS RNA的相对量不超过0.004%。
与早期报告相反,小鼠脾脏中珠蛋白转录本的大幅增加并未伴随着可检测到的珠蛋白mRNA反义RNA水平。