Joint Unit Hospices Civils de Lyon, bioMérieux, Cancer Biomarkers Research Group, Centre Hospitalier Lyon Sud, Lyon, France.
PLoS One. 2012;7(6):e40194. doi: 10.1371/journal.pone.0040194. Epub 2012 Jun 28.
Human endogenous retroviruses (HERVs) are spread throughout the genome and their long terminal repeats (LTRs) constitute a wide collection of putative regulatory sequences. Phylogenetic similarities and the profusion of integration sites, two inherent characteristics of transposable elements, make it difficult to study individual locus expression in a large-scale approach, and historically apart from some placental and testis-regulated elements, it was generally accepted that HERVs are silent due to epigenetic control. Herein, we have introduced a generic method aiming to optimally characterize individual loci associated with 25-mer probes by minimizing cross-hybridization risks. We therefore set up a microarray dedicated to a collection of 5,573 HERVs that can reasonably be assigned to a unique genomic position. We obtained a first view of the HERV transcriptome by using a composite panel of 40 normal and 39 tumor samples. The experiment showed that almost one third of the HERV repertoire is indeed transcribed. The HERV transcriptome follows tropism rules, is sensitive to the state of differentiation and, unexpectedly, seems not to correlate with the age of the HERV families. The probeset definition within the U3 and U5 regions was used to assign a function to some LTRs (i.e. promoter or polyA) and revealed that (i) autonomous active LTRs are broadly subjected to operational determinism (ii) the cellular gene density is substantially higher in the surrounding environment of active LTRs compared to silent LTRs and (iii) the configuration of neighboring cellular genes differs between active and silent LTRs, showing an approximately 8 kb zone upstream of promoter LTRs characterized by a drastic reduction in sense cellular genes. These gathered observations are discussed in terms of virus/host adaptive strategies, and together with the methods and tools developed for this purpose, this work paves the way for further HERV transcriptome projects.
人类内源性逆转录病毒 (HERV) 广泛分布于基因组中,其长末端重复序列 (LTR) 构成了广泛的假定调控序列集合。转座元件所具有的系统发育相似性和丰富的整合位点,使得很难在大规模研究中对单个基因座的表达进行研究,而且历史上除了一些胎盘和睾丸调节元件外,人们普遍认为 HERV 由于表观遗传控制而处于沉默状态。在此,我们引入了一种通用方法,旨在通过最小化交叉杂交风险,最佳地对与 25 个碱基探针相关的单个基因座进行特征描述。因此,我们建立了一个专门针对包含 5573 个 HERV 的集合的微阵列,这些 HERV 可以合理地分配到一个独特的基因组位置。我们使用 40 个正常样本和 39 个肿瘤样本的组合面板获得了 HERV 转录组的第一视图。实验表明,HERV 转录组的近三分之一确实在转录。HERV 转录组遵循趋性规则,对分化状态敏感,而且出人意料的是,似乎与 HERV 家族的年龄无关。在 U3 和 U5 区域内的探针集定义用于为一些 LTR 分配功能(即启动子或 polyA),并揭示(i)自主活性 LTR 广泛受到操作决定论的约束,(ii)与沉默 LTR 相比,活性 LTR 周围的细胞基因密度要高得多,(iii)活性 LTR 和沉默 LTR 之间的相邻细胞基因的配置不同,在启动子 LTR 的上游约 8kb 处表现出明显的 sense 细胞基因减少。这些收集到的观察结果是从病毒/宿主适应策略的角度讨论的,以及为此目的开发的方法和工具一起,为进一步的 HERV 转录组项目铺平了道路。