Mukhopadhyay Debnath, Anant Shrikant, Lee Robert M, Kennedy Susan, Viskochil David, Davidson Nicholas O
Department of Medicine, Washington University Medical School, St. Louis, MO 63110, USA.
Am J Hum Genet. 2002 Jan;70(1):38-50. doi: 10.1086/337952. Epub 2001 Nov 27.
C-->U RNA editing of neurofibromatosis 1 (NF1) mRNA changes an arginine (CGA) to a UGA translational stop codon, predicted to result in translational termination of the edited mRNA. Previous studies demonstrated varying degrees of C-->U RNA editing in peripheral nerve-sheath tumor samples (PNSTs) from patients with NF1, but the basis for this heterogeneity was unexplained. In addition, the role, if any, of apobec-1, the catalytic deaminase that mediates C-->U editing of mammalian apolipoprotein B (apoB) RNA, was unresolved. We have examined these questions in PNSTs from patients with NF1 and demonstrate that a subset (8/34) manifest C-->U editing of RNA. Two distinguishing characteristics were found in the PNSTs that demonstrated editing of NF1 RNA. First, these tumors express apobec-1 mRNA, the first demonstration, in humans, of its expression beyond the luminal gastrointestinal tract. Second, PNSTs with C-->U editing of RNA manifest increased proportions of an alternatively spliced exon, 23A, downstream of the edited base. C-->U editing of RNA in these PNSTs was observed preferentially in transcripts containing exon 23A. These findings were complemented by in vitro studies using synthetic RNA templates incubated in the presence of recombinant apobec-1, which again confirmed preferential editing of transcripts containing exon 23A. Finally, adenovirus-mediated transfection of HepG2 cells revealed induction of editing of apoB RNA, along with preferential editing of NF1 transcripts containing exon 23A. Taken together, the data support the hypothesis that C-->U RNA editing of the NF1 transcript occurs both in a subset of PNSTs and in an alternatively spliced form containing a downstream exon, presumably an optimal configuration for enzymatic deamination by apobec-1.
神经纤维瘤病1型(NF1)信使核糖核酸(mRNA)的C→U核糖核酸编辑将精氨酸(CGA)转变为UGA翻译终止密码子,预计会导致编辑后的mRNA翻译终止。先前的研究表明,来自NF1患者的周围神经鞘瘤样本(PNSTs)存在不同程度的C→U核糖核酸编辑,但这种异质性的原因尚不清楚。此外,介导哺乳动物载脂蛋白B(apoB)核糖核酸C→U编辑的催化脱氨酶载脂蛋白B信使核糖核酸编辑酶1(apobec-1)的作用(如果有的话)尚未明确。我们在来自NF1患者的PNSTs中研究了这些问题,并证明一部分样本(8/34)表现出核糖核酸的C→U编辑。在显示NF1核糖核酸编辑的PNSTs中发现了两个显著特征。首先,这些肿瘤表达apobec-1信使核糖核酸,这是在人类中首次证明其在管腔胃肠道以外的表达。其次,核糖核酸进行C→U编辑的PNSTs显示,在编辑碱基下游的一个可变剪接外显子23A的比例增加。在这些PNSTs中,核糖核酸的C→U编辑优先出现在包含外显子23A的转录本中。使用在重组apobec-1存在下孵育的合成核糖核酸模板进行的体外研究补充了这些发现,该研究再次证实了对包含外显子23A的转录本的优先编辑。最后,腺病毒介导的HepG2细胞转染显示apoB核糖核酸编辑的诱导,以及对包含外显子23A的NF1转录本的优先编辑。综上所述,这些数据支持以下假设:NF1转录本的C→U核糖核酸编辑既发生在一部分PNSTs中,也发生在包含下游外显子的可变剪接形式中,推测这是apobec-1进行酶促脱氨的最佳构型。