Wybier-Franqui J, Tobaly-Tapiero J, Coronel A, Giron M L, Chopin-Robert C, Peries J, Emanoil-Ravier R
UPR A0043, CNRS Rétrovirus et Rétrotransposons des Vertébrés, Hôpital Saint-Louis, Paris, France.
AIDS Res Hum Retroviruses. 1995 Jul;11(7):829-36. doi: 10.1089/aid.1995.11.829.
We have characterized human foamy virus (HFV) proviral DNA and determined HFV expression in a persistent infection model, the Dami megakaryocytic cell line. Molecular studies were performed on parental persistently infected cells (Dami-P), as well as on derived clones (Dami-Cl). We report that in these nonlytic and non-HFV producer cells, viral DNA was found to be integrated into the cellular genome and that the few free proviral forms detected in Dami-P cells were deleted in their 5' LTR. Our molecular analysis indicates the presence of undeleted 5' LTR forms in the integrated provirus within a proviral population mainly composed of deleted forms. In addition, the deletion in the bel1 trans-activator gene, previously described by Saïb et al., was found to be highly predominant. However, in 5-iodo-2'-deoxyuridine treated Dami-Cl cultures, virus production occurred, providing evidence for the presence of complete viral genome. Analysis of HFV expression in Dami-Cl cells, by Northern blot and immunoprecipitation, shows that the most striking difference between cytolytic and persistent HFV infection was the lack of expression of structural viral proteins, in contrast with Bet protein expression, which is maintained. Our data suggest that the Bet protein could be involved in the maintenance of viral persistency and that the persistently infected Dami system provides a suitable model for clarifying its function.
我们已对人泡沫病毒(HFV)前病毒DNA进行了特征分析,并在一种持续感染模型——Dami巨核细胞系中确定了HFV的表达情况。对亲本持续感染细胞(Dami-P)以及衍生克隆(Dami-Cl)进行了分子研究。我们报告称,在这些非裂解性且非HFV产生细胞中,发现病毒DNA整合到了细胞基因组中,并且在Dami-P细胞中检测到的少数游离前病毒形式在其5'长末端重复序列(LTR)中发生了缺失。我们的分子分析表明,在主要由缺失形式组成的前病毒群体中,整合的前病毒中存在未缺失的5' LTR形式。此外,发现此前由赛布等人描述的bel1反式激活基因中的缺失高度占主导。然而,在经5-碘-2'-脱氧尿苷处理的Dami-Cl培养物中,出现了病毒产生,这为完整病毒基因组的存在提供了证据。通过Northern印迹法和免疫沉淀法对Dami-Cl细胞中HFV表达的分析表明,溶细胞性和持续性HFV感染之间最显著的差异在于缺乏结构病毒蛋白的表达,与之形成对比的是,Bet蛋白的表达得以维持。我们的数据表明,Bet蛋白可能参与了病毒持续性的维持,并且持续感染的Dami系统为阐明其功能提供了一个合适的模型。