Probe translation regulation by imaging single protein-mRNA interaction in live cells

Translation converts genetic information encoded in nucleic acids into functional proteins, a process fundamental to cellular function and regulation. However, the mechanics of translation at the single- mRNA level within the native cellular environment remain poorly understood. Recent evidence highlights that mRNAs are not passive conveyors of information but are dynamically regulated, transitioning between translationally active and inactive states in a phenomenon termed "bursting translation." Dysregulation of these processes is linked to diseases such as cancer and neurodegenerative disorders. Despite its importance, the molecular basis of bursting translation, particularly the roles of RNA-binding proteins and mRNA conformation, remains elusive. This project builds upon our pioneering Single-molecule Imaging of Nascent Peptides (SINAPS) technology to elucidate the mechanisms governing translation dynamics of normal and erroneous messages in live cells. Specifically, we will: (1) establish a system to visualize single protein-RNA interactions in live cells, refining single-molecule translation assays and developing tools to correlate protein binding with translation dynamics; (2) investigate how protein factors in ribosome-associated quality control (RQC), such as ZNF598 and ASCC3, interact with substrates to resolve ribosomal collisions; and (3) uncover the role of mRNA looping and initiation factor binding in activating translation. This research will provide a comprehensive understanding of in vivo mRNA translation dynamics and regulation. The novel methodologies and insights generated will pave the way for future studies into translation-related disorders and therapeutic strategies. Project Number: 1R35GM161776-01 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of General Medical Sciences (NIGMS) | Principal Investigator: Bin Wu | Institution: JOHNS HOPKINS UNIVERSITY, BALTIMORE, MD | Award Amount: $426,250 | Activity Code: R35 | Study Section: Special Emphasis Panel[ZRG1 MGG-D (55)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11260711