Pervasive differential splicing in Marek's Disease Virus can discriminate CVI-988 vaccine strain from RB-1B virulent strain in chicken embryonic fibroblasts
Pervasive differential splicing in Marek's Disease Virus can discriminate CVI-988 vaccine strain from RB-1B virulent strain in chicken embryonic fibroblasts
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马立克氏病病毒中普遍存在的差异剪接可以区分鸡胚成纤维细胞中的 CVI-988 疫苗株和 RB-1B 强毒株
DOI：
10.1101/723114
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Sadigh Y
中科院分区：
文献类型：
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作者：
Sadigh Y
马立克氏病是一种严重危害世界家禽健康的疾病。它是由高度传染性的马立克氏病病毒（MDV），一种α疱疹病毒引起的。在这里，我们表明，类似于其他成员的疱疹病毒科，MDV也提出了一个复杂的景观剪接事件，其中大部分是未表征和/或未注释。非常引人注目的是，虽然这一事实的生物学相关性是未知的，我们发现，一些病毒剪接异构体是株特异性的，尽管考虑的菌株，强毒RB-1B和疫苗CVI-988的密切序列相似性。我们通过设计一种检测方法来验证我们的发现，该检测方法基于某些RNA种类的存在来区分由鸡胚胎成纤维细胞中的两种菌株引起的感染。据我们所知，这项研究是有史以来第一次完成这样的结果，强调病毒转录组的全面知识是多么重要，可以了解病毒pathogenicity.ImportanceMarek's disease virus（MDV）导致全球范围内鸡的农业经济重要疾病。虽然商业家禽接种了MDV疫苗，但不可能达到绝育免疫，现有疫苗只能保护鸡只免受疾病症状的影响。接种过疫苗的鸡通常会被强毒株重复感染，将病毒释放到环境中。最有效的MDV疫苗株CVI-988与原型强毒病毒株RB-1B具有>99%的序列同一性。有趣的是，我们的工作表明，尽管它们几乎相同的序列MDV株CVI-988和RB-1B具有显着不同的剪接谱，因此转录组。我们通过实时PCR检测MDV毒株CVI-988表达的、RB-1B毒株转录组中不存在的一些剪接异构体，独立验证了这一发现。这些结果表明，MDV的编码潜力可能比以前认为的要大得多，并表明可能低估了病毒转录组在MDV发病机制和预防中的作用。
Marek’s disease is a major scourge challenging poultry health worldwide. It is caused by the highly contagious Marek’s disease virus (MDV), an alphaherpesvirus. Here we show that, similar to other members of itsHerpesviridaefamily, MDV also presents a complex landscape of splicing events, most of which are uncharacterised and/or not annotated. Quite strikingly, and although the biological relevance of this fact is unknown, we found that a number of viral splicing isoforms are strain-specific despite the close sequence similarity of the strains considered, virulent RB-1B and vaccine CVI-988. We validated our findings by devising an assay that discriminates infections caused by the two strains in chicken embryonic fibroblasts based on the presence of some RNA species. To our knowledge, this study is the first ever to accomplish such a result, emphasizing how important a comprehensive knowledge of the viral transcriptome can be to understand viral pathogenesis.ImportanceMarek’s disease virus (MDV) causes an agro-economically important disease of chickens worldwide. Although commercial poultry are vaccinated against MDV, it is not possible to achieve sterilising immunity, and available vaccines can only protect chickens against the symptoms of the disease. Vaccinated chicken often become superinfected with virulent strains, shedding virus into the environment. The most effective MDV vaccine strain, CVI-988, shares >99% sequence identity with the prototype virulent virus strain RB-1B. Interestingly, our work shows that despite their almost identical sequences MDV strains CVI-988 and RB-1B have significantly different splicing profiles, and hence transcriptomes. We independently validated this discovery by detecting with real-time PCR some splicing isoforms expressed by MDV strain CVI-988 and absent in the transcriptome of the RB-1B strain. These results indicate that the coding potential of MDV might be much larger than previously thought, and suggest a likely underestimation of the role of the viral transcriptome in the pathogenesis and prevention of MDV.