Visualizing and Quantifying mRNA Localization at the Invasive Front of 3D Cancer Spheroids DOI Creative Commons
Konstadinos Moissoglu, Stephen Lockett, Stavroula Mili

et al.

Methods in molecular biology, Journal Year: 2023, Volume and Issue: unknown, P. 263 - 280

Published: Jan. 1, 2023

Abstract Localization of mRNAs at the front migrating cells is a widely used mechanism that functionally supports efficient cell movement. It observed in single on two-dimensional surfaces, as well multicellular three-dimensional (3D) structures and tissue vivo. 3D cultures can reveal how topology extracellular matrix cell-cell contacts influence subcellular mRNA distributions. Here we describe method for imaging an inducible system collective cancer invasion. MDA-MB-231 spheroids are embedded Matrigel, induced to invade, processed image with single-molecule sensitivity. An analysis algorithm quantify compare distributions invasive leader cells. The approach be easily adapted applied analyze RNA additional settings where polarize along linear axis.

Language: Английский

The role of midbody-associated mRNAs in regulating abscission DOI Creative Commons
Trey Farmer, Katherine F. Vaeth,

Ke-Jun Han

et al.

The Journal of Cell Biology, Journal Year: 2023, Volume and Issue: 222(12)

Published: Nov. 3, 2023

Midbodies function during telophase to regulate the abscission step of cytokinesis. Until recently, it was thought that abscission-regulating proteins, such as ESCRT-III complex subunits, accumulate at MB by directly or indirectly binding resident protein, CEP55. However, recent studies have shown depletion CEP55 does not fully block targeting MB. Here, we show MBs contain mRNAs and these MB-associated can be locally translated, resulting in accumulation proteins. We demonstrate localized translation CHMP4B is required for its site 3′ UTR-dependent mRNA successful completion Finally, identify regulatory cis-elements within RNAs are necessary sufficient trafficking propose a novel method regulating cytokinesis selective mRNAs.

Language: Английский

Citations

11
A KIF1C-CNBP motor-adaptor complex for trafficking mRNAs to cell protrusions DOI Creative Commons
Konstadinos Moissoglu, Tianhong Wang, Alexander N. Gasparski

et al.

Cell Reports, Journal Year: 2025, Volume and Issue: 44(3), P. 115346 - 115346

Published: Feb. 20, 2025

Language: Английский

Citations

0
RAB13 mRNA compartmentalisation spatially orients tissue morphogenesis DOI Creative Commons
Guilherme Costa, Joshua J Bradbury, Nawseen Tarannum

et al.

The EMBO Journal, Journal Year: 2020, Volume and Issue: 39(21)

Published: Sept. 18, 2020

Polarised targeting of diverse mRNAs to cellular protrusions is a hallmark cell migration. Although widespread phenomenon, definitive functions for endogenous targeted and their relevance modulation in vivo tissue dynamics remain elusive. Here, using single-molecule analysis, gene editing zebrafish live-cell imaging, we report that mRNA polarisation acts as molecular compass orients motile polarity spatially directs movement. Clustering protrusion-derived RNAseq datasets defined core 192-nt localisation element underpinning precise sites filopodia formation. Such the small GTPase RAB13 generated tight spatial coupling localisation, translation protein activity, achieving subcellular compartmentalisation function create polarised domain extension. Consequently, genomic excision this perturbation targeting-but not translation-depolarised endothelial cells induced mispatterning blood vessels zebrafish. Hence, polarisation, expression, primary determinant site action, preventing ectopic functionality at inappropriate loci orienting morphogenesis.

Language: Английский

Citations

31
RNA‐binding proteins and translation control in angiogenesis DOI Open Access
Madeleine Smith, Guilherme Costa

FEBS Journal, Journal Year: 2021, Volume and Issue: 289(24), P. 7788 - 7809

Published: Nov. 19, 2021

Tissue vascularization through the process of angiogenesis ensures adequate oxygen and nutrient supply during development regeneration. The complex morphogenetic events involved in new blood vessel formation are orchestrated by a tightly regulated crosstalk between extra intracellular factors. In this context, RNA‐binding protein (RBP) activity translation play fundamental roles cellular responses triggered particular environmental cues. A solid body work has demonstrated that key RBPs (such as HuR, TIS11 proteins, hnRNPs, NF90, QKIs YB1) implicated both physiological pathological angiogenesis. These critical for metabolism messenger (m)RNAs encoding angiogenic modulators and, importantly, strong evidence suggests RBP–mRNA interactions can be altered disease. Lesser known, but not less important, mechanistic aspects synthesis also regulate generation vessels. review, we outline findings demonstrating implications RBP‐mediated RNA regulation control Furthermore, highlight how these mechanisms post‐transcriptional gene expression have led to promising therapeutic strategies aimed at targeting undesired formation.

Language: Английский

Citations

21
The scaffolding protein AKAP12 regulates mRNA localization and translation DOI Creative Commons
Madeleine Smith,

Parisa Naeli,

Seyed Mehdi Jafarnejad

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2024, Volume and Issue: 121(18)

Published: April 23, 2024

Regulation of subcellular messenger (m)RNA localization is a fundamental biological mechanism, which adds spatial dimension to the diverse layers post-transcriptional control gene expression. The cellular compartment in mRNAs are located may define distinct aspects encoded proteins, ranging from production rate and complex formation localized activity. Despite detailed roles that have emerged over past decades, identity factors anchoring domains remains ill-defined. Here, we used an unbiased method profile RNA-bound proteome migrating endothelial cells (ECs) discovered plasma membrane (PM)–associated scaffolding protein A-kinase anchor (AKAP)12 interacts with various mRNAs, including transcripts encoding kinases Actin remodeling In particular, AKAP12 targets transcript coding for kinase Abelson Tyrosine-Protein Kinase 2 (ABL2), found be necessary adequate filopodia angiogenic sprouting. Moreover, demonstrate ABL2 mRNA PM show absence AKAP12, translation efficiency reduced. Altogether, our work identified unique function sheds light into mechanisms

Language: Английский

Citations

2
RNA localization in confined cells depends on cellular mechanical activity and contributes to confined migration DOI Creative Commons

Rebecca A. Moriarty,

Stavroula Mili, Kimberly M. Stroka

et al.

iScience, Journal Year: 2022, Volume and Issue: 25(2), P. 103845 - 103845

Published: Feb. 1, 2022

Cancer cells experience mechanical confining forces during metastasis and, consequently, can alter their migratory mechanisms. Localization of numerous mRNAs to cell protrusions contributes polarization and migration is controlled by proteins that bind RNA and/or cytoskeletal elements, such as the adenomatous polyposis coli (APC). Here, we demonstrate peripheral localization APC-dependent RNAs in within confined microchannels type dependent. This varying phenotype determined levels a detyrosinated tubulin network. We show this network regulated mechanoactivity with mechanosensitive ion channels increased myosin II activity direct RAB13 RNA. Through specific mislocalization RNA, migration. Our results indicate cell's determines its ability peripherally target utilize them for movement confinement.

Language: Английский

Citations

10
A KIF1C-CNBP motor-adaptor complex for trafficking mRNAs to cell protrusions DOI
Konstadinos Moissoglu, Tianhong Wang, Alexander N. Gasparski

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 27, 2024

Summary mRNA localization to subcellular compartments is a widely used mechanism that functionally contributes numerous processes. targeting can be achieved upon recognition of RNA cargo by molecular motors. However, our understanding how this accomplished limited, especially in higher organisms. We focus on pathway targets mRNAs peripheral protrusions mammalian cells and important for cell migration. Trafficking occurs through active transport microtubules, mediated the KIF1C kinesin. Here, we identify RNA-binding protein CNBP, as factor required protrusions. CNBP binds directly GA-rich sequences 3’UTR protrusion targeted mRNAs. also interacts with recruitment their trafficking microtubules periphery. This work provides reveals motor-adaptor complex

Language: Английский

Citations

1
Insights into the regulation of mRNA translation by scaffolding proteins DOI Creative Commons
Madeleine Smith, Guilherme Costa

Biochemical Society Transactions, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 6, 2024

Regionalisation of molecular mechanisms allows cells to fine-tune their responses dynamic environments. In this context, scaffolds are well-known mediators localised protein activity. These phenomenal proteins act as docking sites where pathway components brought together ensure efficient and reliable flow information within the cell. Although mostly understood hubs for signalling communication, some have also been studied regulators mRNA translation. Here, we provide a brief overview work unravelling how scaffolding facilitate cross-talk between two processes. Firstly, examine activity AKAP1 AKAP12, that not only capacity anchor mRNAs membranes but can regulate synthesis. Next, review studies uncovered ribosome-associated RACK1 orchestrates translation initiation. We discuss evidence pointing Ezrin LASP1 early stages. end, conclude with open questions propose future directions will bring new insights into regulation by proteins.

Language: Английский

Citations

1
Cell Migration in Three Dimensions DOI
Coert Margadant

Methods in molecular biology, Journal Year: 2023, Volume and Issue: unknown, P. 1 - 14

Published: Jan. 1, 2023

Language: Английский

Citations

3
The kinesin KIF1C transports APC-dependent mRNAs to cell protrusions DOI Open Access
Xavier Pichon, Konstadinos Moissoglu,

Emeline Coleno

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2020, Volume and Issue: unknown

Published: Nov. 30, 2020

Abstract RNA localization and local translation are important for numerous cellular functions. In mammals, a class of mRNAs localize to cytoplasmic protrusions in an APC-dependent manner, with roles during cell migration. Here, we investigated this mechanism. We found that the KIF1C motor interacts is required their localization. Live imaging revealed rapid, active transport single over long distances requires both microtubules KIF1C. Two color directly showed transported by motors, 3’UTR being sufficient trigger KIF1C-dependent Moreover, remained associated peripheral, multimeric clusters was formation. These results reveal pathway mammalian cells, which has dual role transporting RNAs promoting clustering within protrusions. Interestingly, also transports its own mRNA suggesting possible feedback loop acting at level transport.

Language: Английский

Citations

4