Epitranscriptomic Regulation of Alphavirus Replication and Transmission

摘要 甲型病毒是全球公共卫生面临的一个不断出现的挑战。这方面的例证是 基孔肯雅病毒从非洲通过南亚传播到美洲。另外，这段过去 夏季东部马脑炎病毒感染和死亡人数有所增加 美国。需要更深入地了解有效传播和复制的决定因素 确定传播干预的目标。近年来，已经很明显的是，对 病毒RNA影响其功能，并影响感染的结局。拟议的研究扩展了已发表的和 初步观察表明病毒基因组RNA具有特殊的表位转录修饰 传动效率的确定。这一建议调查了寄主物种的新假设 包装的病毒基因组RNA的特定修饰会影响颗粒确定感染的能力 在下一个宿主细胞中，甲型病毒必须在 蚊子和脊椎动物的宿主。为了检验这一点，拟议的研究将：(I)采用多重排序 确定产生的颗粒物基因组中胞嘧啶残基甲基化状态的技术 脊椎动物和蚊子细胞；(Ii)检查破坏病毒甲基化的生物学后果 通过改变甲基转移酶和/或发现经常被修改的突变残基来改变基因组。这个 这项工作的结果将阐明病毒RNA胞嘧啶甲基化的亲病毒和抗病毒作用，并为 为进一步研究宿主特异性表位转录修饰及其在病毒中的作用奠定基础 复制和传播。

Summary Alphaviruses represent a continuingly emerging challenge to global public health. This is exemplified by the spread of chikungunya virus from Africa through southern Asia and into the Americas. Additionally, this past summer has seen an increase in the number of eastern equine encephalitis virus infections and deaths in the United States. A deeper understanding of the determinants of efficient transmission and replication is needed to identify targets for transmission interventions. In recent years it has become apparent that modification of viral RNA impacts it’s function, and the outcome of infection. The proposed research extends on published and preliminary observations that implicate a particular epitranscriptomic modification of the viral genomic RNA in the determination of transmission efficiency. This proposal investigates the novel hypothesis that host species specific modification of the packaged viral genomic RNA impacts the ability of the particle to establish infection in the next host cell, a prospect of significant consequence for alphaviruses that obligatorily cycle between mosquito and vertebrate hosts. To examine this the proposed research will: (i) employ multiple sequencing technologies to determine the methylation state of cytosine residues in the genome of particles produced in vertebrate and mosquito cells; (ii) examine the biological consequence of disrupting the methylation of the viral genome through altering the methyltransferase and/or mutating residues found to be frequently modified. The results of this work will elucidate the pro- and anti-viral effects of cytosine methylation of viral RNA, and lay the ground work for further investigation of host-specific epitranscriptomic modifications and their role in virus replication and transmission.