Epitranscriptomic Regulation of Alphavirus Replication and Transmission

总结 甲病毒是对全球公共卫生的持续出现的挑战。这一点的例证是， 基孔肯雅病毒从非洲经南亚传播到美洲。此外，过去 夏季，东部马脑炎病毒感染和死亡人数有所增加， 美国的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，人们发现， 病毒RNA影响它的功能和感染的结果。拟议的研究扩展到已发表的和 初步的观察结果表明，一个特定的表观修饰的病毒基因组RNA， 传输效率的确定。这项提议调查了一个新的假设，即宿主物种 包装的病毒基因组RNA的特异性修饰影响颗粒建立感染的能力 在下一个宿主细胞中，甲病毒的显著后果的前景是， 蚊子和脊椎动物宿主。为了检验这一点，拟议的研究将：（i）采用多重测序 技术，以确定甲基化状态的胞嘧啶残基的基因组中产生的颗粒， 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/或突变发现经常被修饰的残基来改变基因组。的 这项工作的结果将阐明病毒RNA的胞嘧啶甲基化的促病毒和抗病毒作用，并奠定 进一步研究宿主特异性表位转录组修饰及其在病毒中的作用的基础工作 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.