Centre for Chromosome Biology, School of Natural Sciences, National University of Ireland, Galway, Galway, Ireland.
J Virol. 2011 Jul;85(14):6930-40. doi: 10.1128/JVI.00547-11. Epub 2011 May 4.
Small noncoding RNAs regulate a variety of cellular processes, including genomic imprinting, chromatin remodeling, replication, transcription, and translation. Here, we report small replication-regulating RNAs (srRNAs) that specifically inhibit DNA replication of the human BK polyomavirus (BKV) in vitro and in vivo. srRNAs from FM3A murine mammary tumor cells were enriched by DNA replication assay-guided fractionation and hybridization to the BKV noncoding control region (NCCR) and synthesized as cDNAs. Selective mutagenesis of the cDNA sequences and their putative targets suggests that the inhibition of BKV DNA replication is mediated by srRNAs binding to the viral NCCR, hindering early steps in the initiation of DNA replication. Ectopic expression of srRNAs in human cells inhibited BKV DNA replication in vivo. Additional srRNAs were designed and synthesized that specifically inhibit simian virus 40 (SV40) DNA replication in vitro. These observations point to novel mechanisms for regulating DNA replication and suggest the design of synthetic agents for inhibiting replication of polyomaviruses and possibly other viruses.
小非编码 RNA 调控多种细胞过程，包括基因组印记、染色质重塑、复制、转录和翻译。在这里，我们报告了小复制调控 RNA（srRNA），它可以特异性抑制人 BK 多瘤病毒（BKV）在体外和体内的 DNA 复制。通过 DNA 复制测定指导的分级分离和与 BKV 非编码调控区（NCCR）杂交富集 FM3A 鼠乳腺肿瘤细胞中的 srRNA，并将其合成 cDNA。对 cDNA 序列及其潜在靶标的选择性诱变表明，BKV DNA 复制的抑制是通过 srRNA 与病毒 NCCR 结合介导的，从而阻碍了 DNA 复制起始的早期步骤。srRNA 在人细胞中的异位表达抑制了体内的 BKV DNA 复制。设计并合成了其他 srRNA，可特异性抑制猿猴病毒 40（SV40）在体外的 DNA 复制。这些观察结果指出了调节 DNA 复制的新机制，并为设计抑制多瘤病毒和可能其他病毒复制的合成剂提供了思路。
