Structural studies with regulatory RNAs

点击翻译按钮获取中文摘要

Collectively, our Meg3 study answers basic questions regarding structure and activity of this lncRNA, providing a template for functional and mechanistic studies of other lRNAs of this family. These findings also lay a framework for efficiently harnessing the tumor suppressive property of the RNA for potential anti-cancer therapies. Meg3 RNA is by far the best-characterized tumor suppressor lncRNA. In various cancer cell lines and clinical samples, Meg3 is either not expressed or is expressed at low levels, and exogenous expression of Meg3 RNA in such cells can slow growth and induce apoptosis. Targeted restoration or overexpression of Meg3 RNA in affected cells, therefore, offers a promising avenue for cancer treatment. Our structural probing data for KSHV PAN lncRNA pave the way for a comprehensive reconstruction of the protein-lncRNA interaction network in various biological contexts. PAN RNA is a highly multifunctional viral transcript that mediates its effects through locally structured RNA segments and interactions with several viral and host proteins. Additional studies of PAN and other KSHV lncRNAs will further elucidate the complex molecular biology of this important pathogen. NMR studies with hepatitis B virus (HBV) have revealed novel mechanistic insights into the packaging element, epsilon, as it relates to initiation of DNA synthesis, providing additional avenues for therapeutic intervention. In collaboration with the NIAID Integrated Research Facility, we have determined the structure of an Ebola virus minigenome. Herein, we demonstrated that the 5 prime NCR of the EBOV RNA genome forms complex secondary and tertiary domains that interact with a host protein and are essential for virus replication. Our data indicate that host protein HSPA8 interacts with a specific RNA motif that forms the right side of a stem-loop located in the EBOV trailer (nt 28-37). This interaction is essential for EBOV minigenome replication and recovery of full-length infectious virus from cDNA. These studies lay a foundation for development of ligands that target the Ebola virus genome. Work on this project concluded in FY21.