2023
DOI: 10.1038/s41592-023-02101-9
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System-wide analysis of RNA and protein subcellular localization dynamics

Eneko Villanueva,
Tom Smith,
Mariavittoria Pizzinga
et al.

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 … Show more

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Cited by 15 publications
(4 citation statements)
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“…Protein functions are determined by their subcellular localization within various organelles, where they function in different environments (Pan et al, 2021, Villanueva et al, 2024. While examining locations of up-regulated proteins in both the susceptible and wild tomato plants, the predominant subcellular locations were the intracellular regions, followed by cellular anatomical entities.…”
Section: Discussionmentioning
confidence: 99%
“…Protein functions are determined by their subcellular localization within various organelles, where they function in different environments (Pan et al, 2021, Villanueva et al, 2024. While examining locations of up-regulated proteins in both the susceptible and wild tomato plants, the predominant subcellular locations were the intracellular regions, followed by cellular anatomical entities.…”
Section: Discussionmentioning
confidence: 99%
“…These methods often rely on cellular fractionation approaches or proximity labeling techniques as prerequisite steps . A diverse range of subcellular fractionation techniques, such as detergent-based fractionation, centrifugation-sedimentation-based, density centrifugation-based methods, electrophoresis, and affinity purification, are available to support MS-based workflows. Furthermore, organellar profiling methods have been extensively applied to cell-wide and single organelles.…”
Section: Subcellular Proteomics Methodsmentioning
confidence: 99%
“…A prerequisite for multiorganelle protein mapping is physical cell disruption to release intact organelles (Table 1) for subcellular compartment enrichment. Further, intact organelles are enriched by density centrifugation 37 or differential centrifugation 21 methods. Different organelles are enriched in density-dependent methods, but their distribution is not absolute and can significantly overlap.…”
Section: Mass Spectrometry-based Methodsmentioning
confidence: 99%
“…System-wide studies of RNA subcellular localization (e.g., mRNA [177]) have also paved the way for a more comprehensive analysis of the cellular dynamics [178,179], as proteins are usually transcribed by RNA molecules. Moreover, except for RNA transcripts for protein, other RNAs, like long noncoding RNAs (lncRNAs), may also be involved in many biological functions [180].…”
Section: Future Directionsmentioning
confidence: 99%