RNP transport in cell biology: the long and winding road

Eliscovich, Carolina; Singer, Robert H.

doi:10.1016/j.ceb.2017.02.008

Cited by 69 publications

(68 citation statements)

References 96 publications

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“…We next set out to determine the molecular mechanism of RP-mRNAs localization to protrusions. Localization of mRNAs is driven by specific RBPs which bind to and mediate transport or subcellular anchoring of their target transcripts (Eliscovich and Singer, 2017). We 145 therefore hypothesized that specific protrusion localized RBPs must be interacting with and localizing RP-mRNAs to protrusions.…”

Section: Depletion Of Larp Proteins Reveals a Role For Larp6 In Rp-mrmentioning

confidence: 99%

Subcellular mRNA localization regulates ribosome biogenesis in migrating cells

Dermit

Dodel

Lee

et al. 2019

Preprint

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Translation of Ribosomal Protein coding mRNAs (RP-mRNAs) constitutes a key step in regulation of ribosome biogenesis, but the mechanisms which modulate RP-mRNAs 20 translation under various cellular and environmental conditions are poorly understood. Here we show that the subcellular localization of RP-mRNAs acts as a key regulator of their translation in migrating cells. As cells migrate into their surroundings, RP-mRNAs localize to the actin-rich protrusions at the front the cells. This localization is mediated by La related protein-6 (LARP6), an RNA Binding Protein that is enriched in protrusions. Protrusions act as 25 hotspots of translation for localized RP-mRNAs, resulting in enhancement of RP synthesis, upregulation of ribosome biogenesis, and increased overall protein synthesis of migrating cells in a LARP6 dependent manner. In human breast carcinomas, Epithelial to Mesenchymal Transition (EMT) upregulates LARP6 expression to enhance protein synthesis, and can be targeted using a small molecule inhibitor that interferes with LARP6 RNA binding. Our findings 30 reveal LARP6 mediated mRNA localization as a key regulator of ribosome biogenesis during cell-migration, and demonstrate a role for this process in cancer progression downstream of EMT.35 65 mRNAs to promote their enrichment in protrusions. Protrusions are also highly enriched in translation initiation and elongation factors, acting as hotspots for translation of localized RP-mRNAs. LARP6 dependent localization of RP-mRNAs results in upregulation of RP levels, leading to enhancement of ribosome biogenesis and global protein synthesis in migrating cells. In human breast carcinomas, higher LARP6 protein expression is associated with the 70 invasive mesenchymal-like subtypes. EMT induces LARP6 protein expression, which acts to promote malignant growth and invasion. Crucially, LARP6 regulation of RP-mRNAs can be therapeutically targeted using small molecule inhibitors which specifically interfere with its RNA binding. Collectively, our findings reveal a new mechanism that governs ribosome biogenesis in mesenchymal-like migratory cells via subcellular localization of RP-mRNAs, and 75 demonstrate a therapeutically targetable role for this process in cancer downstream of EMT. Results 80 RP-mRNAs localize to protrusions of all migratory cellsWe utilized a micro-porous transwell filter based method (Mardakheh et al., 2015;Mili et al., 2008) to assess if RP-mRNAs localization to actin-rich protrusions was a conserved feature of all migratory cells. We modified the procedure to allow cells to adhere to the top of the filter first, followed by synchronized induction of protrusion formation through the pores (Figure 85 1A). Importantly, the small (3µm) size of the pores enables protrusions to form but prevents the cell bodies from passing through, thus resulting in separation of the protrusions and the cell bodies on opposite sides of the filter, which can be independently imaged or purified for multi-omics analysis ( Figure 1A). Using this method, we profiled ...

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Section: Depletion Of Larp Proteins Reveals a Role For Larp6 In Rp-mrmentioning

confidence: 99%

Subcellular mRNA localization regulates ribosome biogenesis in migrating cells

Dermit

Dodel

Lee

et al. 2019

Preprint

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“…Interestingly, different cell types such as neurons with protrusions exceeding hundreds of µm, and compact cell types such as fibroblasts, utilize different mechanisms to distribute β-actin mRNA 20 . The diversity of mRNA dynamics highlights the need to investigate mRNA movement in different cell types and interrogate the complex network of mRNA binding proteins and their role for distributing mRNAs within cells 21 .…”

Section: Introductionmentioning

confidence: 99%

Detection and quantification of single mRNA dynamics with the Riboglow fluorescent RNA tag

Braselmann

Stasevich

Lyon

et al. 2019

Preprint

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Labeling and tracking biomolecules with fluorescent probes on the single molecule level enables quantitative insights into their dynamics in living cells. We previously developed Riboglow, a platform to label RNAs in live mammalian cells, consisting of a short RNA tag and a small organic probe that increases fluorescence upon binding RNA. Here, we demonstrate that Riboglow is capable of detecting and tracking single RNA molecules. We benchmark RNA tracking by comparing results with the established MS2 RNA tagging system. To demonstrate versatility of Riboglow, we assay translation on the single molecule level, where the translated mRNA is tagged with Riboglow and the nascent polypeptide is labeled with a fluorescent antibody. The growing effort to investigate RNA biology on the single molecule level requires sophisticated and diverse fluorescent probes for multiplexed, multi-color labeling of biomolecules of interest, and we present Riboglow as a new member in this toolbox.

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“…Studies from several model transcripts have uncovered general principles that dictate how mRNAs become localized within cells. Conserved cis ‐acting sequences, usually within the mRNA 3′‐untranslated region (3′UTR), are recognized by trans ‐acting RNA‐binding proteins (RBPs) to regulate mRNA transport, translation, stability and degradation . Each RBP can interact with few or many target mRNAs on the basis of primary sequence and/or secondary structure.…”

Section: Introductionmentioning

confidence: 99%

“…RNA-binding proteins (RBPs) to regulate mRNA transport, translation, stability and degradation. 2 Each RBP can interact with few or many target mRNAs on the basis of primary sequence and/or secondary structure. Most localized mRNAs interact with multiple RBPs throughout their lifetime, and the localization of multiple mRNAs to the same subcellular domain is often regulated by the same RBPs.…”

mentioning

confidence: 99%

“…Most localized mRNAs interact with multiple RBPs throughout their lifetime, and the localization of multiple mRNAs to the same subcellular domain is often regulated by the same RBPs. 2 One key function of RBPs is to coordinate the active transport of mRNAs to subcellular targets via molecular motors that move along cytoskeletal networks (reviewed by Gagnon and Mowry 3 and Bullock 4 ). All three families of molecular motors-myosins, kinesins and dynein-are known to transport mRNAs.…”

mentioning

confidence: 99%

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RNA localization regulates diverse and dynamic cellular processes

Ryder

Lerit

2018

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At the nexus of specialized cellular responses are localized enrichments of protein activity. The localization of messenger RNA (mRNA) coupled with translational control often plays a crucial role in the generation of protein concentrations at defined subcellular domains. Although mRNA localization is classically associated with large specialized cells, such as neurons and embryos, RNA localization is a highly conserved paradigm of post-transcriptional regulation observed in diverse cellular contexts. Functions of localized mRNAs extend far beyond the well-studied examples of neuronal polarization and developmental patterning. Since the initial discovery of the intracellular localization of cytoskeletal mRNAs within migrating cells, hundreds of mRNAs are now known to be enriched at specific organelles where they contribute to cell function. In this short review, we discuss basic principles regulating RNA localization and consider the contribution of localized mRNA to several essential cellular behaviors. We consider RNA localization as a mechanism with widespread implications for cellular function.

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RNP transport in cell biology: the long and winding road

Cited by 69 publications

References 96 publications

Subcellular mRNA localization regulates ribosome biogenesis in migrating cells

Subcellular mRNA localization regulates ribosome biogenesis in migrating cells

Detection and quantification of single mRNA dynamics with the Riboglow fluorescent RNA tag

RNA localization regulates diverse and dynamic cellular processes

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