학술논문

System-wide analysis of RNA and protein subcellular localization dynamics
Document Type
Original Paper
Source
Nature Methods: Techniques for life scientists and chemists. 21(1):60-71
Subject
Language
English
ISSN
1548-7091
1548-7105
Abstract
Although the subcellular dynamics of RNA and proteins are key determinants of cell homeostasis, their characterization is still challenging. Here we present an integrative framework to simultaneously interrogate the dynamics of the transcriptome and proteome at subcellular resolution by combining two methods: localization of RNA (LoRNA) and a streamlined density-based localization of proteins by isotope tagging (dLOPIT) to map RNA and protein to organelles (nucleus, endoplasmic reticulum and mitochondria) and membraneless compartments (cytosol, nucleolus and cytosolic granules). Interrogating all RNA subcellular locations at once enables system-wide quantification of the proportional distribution of RNA. We obtain a cell-wide overview of localization dynamics for 31,839 transcripts and 5,314 proteins during the unfolded protein response, revealing that endoplasmic reticulum-localized transcripts are more efficiently recruited to cytosolic granules than cytosolic RNAs, and that the translation initiation factor eIF3d is key to sustaining cytoskeletal function. Overall, we provide the most comprehensive overview so far of RNA and protein subcellular localization dynamics.
An integrative framework to simultaneously interrogate the dynamics of the transcriptome and proteome at subcellular resolution that combines two methods, localization of RNA (LoRNA) and a streamlined density-based localization of proteins by isotope tagging (dLOPIT).