The role of asymmetric protein dimethylation during the human papillomavirus lifecycle

不对称蛋白质二甲基化在人乳头瘤病毒生命周期中的作用

• 批准号: 10518761
• 负责人: Koenraad Van Doorslaer
• 金额: $ 38.05万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-07 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10518761
• 关键词: 3-Dimensional; Address; Affect; Alternative Splicing; Anogenital cancer; Biological Assay; Biology; Cell physiology; Cells; Cervical; Chromosomes; Clinical Trials; Complex; DNA Damage; Data; Deposition; Development; Environment; Enzymes; Epithelial; Event; Exons; Genome; Genomic approach; Head and Neck Cancer; Head and neck structure; Histones; Human; Human Papilloma Virus-Related Malignant Neoplasm; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 18; Incidence; Infection; Introns; Knowledge; Lead; Length; Link; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mediating; Messenger RNA; Methylation; Methyltransferase; Modification; Papillomavirus; Pattern; Phase; Phenotype; Play; Polyadenylation; Primary Infection; Production; Protein Inhibition; Protein Overexpression; Protein-Arginine N-Methyltransferase; Proteins; RNA; RNA Splicing; RNA methylation; Regulation; Risk Factors; Role; Sexually Transmitted Diseases; Site; Testing; Tissues; Transcript; Viral; Viral Genes; Viral Genome; Viral Proteins; Virus; Virus Diseases; Virus Replication; Work; acute infection; chronic infection; druggable target; human papilloma virus oncogene; keratinocyte differentiation; novel therapeutics; single-cell RNA sequencing; spatiotemporal; tool; transcriptome; viral DNA

Infections with human papillomaviruses (HPVs) are the most common sexually transmitted infection in the US. In addition to causing cervical and other anogenital cancers and a rising incidence of head and neck cancer, HPVs are responsible for an estimated 5% of cancers worldwide. Importantly, persistent infection, and not an acute infection is the primary risk factor for (cervical) cancer development. Thus, understanding the virus-host interplay that promotes or restrict viral persistence has important implications for HPV biology and human cancers. Until recently, it has been challenging to study the immediate early events of the HPV lifecycle following infection and how these events contribute to initial genome amplification during the establishment phase and long-term viral persistence. We used a single-cell genomics approach to identify cellular factors involved in viral infection and persistence. Our preliminary data identify protein arginine N-methyltransferase 1 (PRMT1) as an important factor upon viral infection of primary human cervical cells. PRMT1 regulates several cellular functions that may be relevant to the HPV lifecycle. We demonstrate that PRMT1 inhibition leads to a highly dysregulated splicing pattern of the viral genes. Mechanistically, we identify a dramatic increase in m6A modifications of viral mRNA, specifically in introns. These data implicate RBM15, a regulatory subunit of the m6A methyltransferase complex. We hypothesize that PRMT1 regulates RBM15 controlled m6a deposition on viral mRNA to temporally regulate alternative splicing throughout viral infection. We will (1) Determine the importance of asymmetric protein dimethylation throughout the viral lifecycle. (2) Determine the role for RBM15 in the PRMT1 mediated regulation of m6a methylation of viral mRNA. (3) Determine the effects of m6a methylation on viral mRNA splicing. PRMT1 inhibition leads to dysregulated viral splicing. This proposal addresses a critical gap in our knowledge relating to how HPV splicing is regulated for optimal infection, establishment, and long-term persistence. We identify PRMT1 as a regulator of viral splicing and hypothesize that this occurs through targeted m6A deposition on viral introns. Since PRMT1 is an established druggable target, these results may lead to novel therapeutics for HPV-induced cancers.

人乳头瘤病毒 (HPV) 感染是美国最常见的性传播感染。除了导致宫颈癌和其他肛门生殖器癌症以及头颈癌发病率上升之外，HPV 还导致全球约 5% 的癌症。重要的是，持续感染而不是急性感染是（宫颈）癌发展的主要危险因素。因此，了解促进或限制病毒持久性的病毒-宿主相互作用对于 HPV 生物学和人类癌症具有重要意义。直到最近，研究感染后 HPV 生命周期的早期事件以及这些事件如何促进建立阶段的初始基因组扩增和长期病毒持续存在一直是一项挑战。我们使用单细胞基因组学方法来识别与病毒感染和持久性有关的细胞因素。我们的初步数据表明，蛋白精氨酸 N-甲基转移酶 1 (PRMT1) 是原代人宫颈细胞病毒感染的重要因素。 PRMT1 调节可能与 HPV 生命周期相关的多种细胞功能。我们证明 PRMT1 抑制会导致病毒基因的剪接模式高度失调。从机制上讲，我们发现病毒 mRNA 的 m6A 修饰显着增加，特别是在内含子中。这些数据暗示 RBM15，m6A 甲基转移酶复合物的调节亚基。我们假设 PRMT1 调节 RBM15 控制的 m6a 在病毒 mRNA 上的沉积，从而在整个病毒感染过程中暂时调节选择性剪接。我们将 (1) 确定不对称蛋白质二甲基化在整个病毒生命周期中的重要性。 (2)确定RBM15在PRMT1介导的病毒mRNA m6a甲基化调节中的作用。 (3)确定m6a甲基化对病毒mRNA剪接的影响。 PRMT1 抑制会导致病毒剪接失调。该提案解决了我们在如何调节 HPV 剪接以实现最佳感染、建立和长期持续性方面的知识中的一个关键空白。我们将 PRMT1 确定为病毒剪接的调节因子，并假设这是通过病毒内含子上的靶向 m6A 沉积而发生的。由于 PRMT1 是一个已确定的药物靶点，这些结果可能会带来针对 HPV 诱导的癌症的新疗法。