In vivo single-particle imaging of nuclear mRNA export in budding yeast demonstrates an essential role for Mex67p

Smith, Carlas; Lari, Abdolaziz Rastegar; Derrer, Carina Patrizia; Ouwehand, Arlieneke; Rossouw, Ammeret; Huisman, Maximiliaan; Dange, Thomas; Hopman, Mark; Joseph, Aviva; Zenklusen, Daniel; Weis, Karsten; Grünwald, David; Montpetit, Ben

doi:10.1083/jcb.201503135

Cited by 67 publications

(76 citation statements)

References 61 publications

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“…Our molecular architecture is fully consistent with proposed mRNP remodeling models (Folkmann et al, 2011; Montpetit et al, 2011), as well as with the observation that cytoplasmic release - but not translocation - is a rate limiting step during mRNA export (Oeffinger and Zenklusen, 2012). When translated into the overall NPC architecture, the presence of eight remodeling hubs surrounding the central channel ensures a highly efficient system consistent with the fast mRNA export rates observed in vivo (Grunwald and Singer, 2010; Mor et al, 2010; Smith et al, 2015). Other types of ribonucleoproteins are also actively exported through the NPC, using pathways and components that largely overlap with those of mRNA export (Nerurkar et al, 2015).…”

Section: Discussionmentioning

confidence: 61%

Structure and Function of the Nuclear Pore Complex Cytoplasmic mRNA Export Platform

Fernández-Martı́nez

Kim

Shi

et al. 2016

Cell

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151

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Summary The last steps in mRNA export and remodeling are performed by the Nup82 complex, a large conserved assembly at the cytoplasmic face of the nuclear pore complex (NPC). By integrating diverse structural data, we have determined the molecular architecture of the native Nup82 complex at subnanometer precision. The complex consists of two compositionally identical multiprotein subunits that adopt different configurations. The Nup82 complex fits into the NPC through the outer ring Nup84 complex. Our map shows that this entire 14 MDa Nup82-Nup84 complex assembly positions the cytoplasmic mRNA export factor docking sites and mRNP remodeling machinery right over the NPC's central channel, rather than on distal cytoplasmic filaments as previously supposed. We suggest that this configuration efficiently captures and remodels exporting mRNP particles immediately upon reaching the cytoplasmic side of the NPC.

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Section: Discussionmentioning

confidence: 61%

Structure and Function of the Nuclear Pore Complex Cytoplasmic mRNA Export Platform

Fernández-Martı́nez

Kim

Shi

et al. 2016

Cell

Self Cite

151

218

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“…Once in the cytoplasm, the mRNPs are confined near the nuclear envelope. This suggests that mRNPs, once exported, remain with multiple NPCs at their cytoplasmic face; most likely, the mRNP composition is remodeled for future events in the cytoplasm (133). Visualization of the interaction between specific RNA-binding proteins (RBPs) and single-mRNA molecules is essential to understanding the metabolism of the mRNA in the cytoplasm.…”

Section: Cotranscriptional and Post-transcriptional Regulation Of Genmentioning

confidence: 99%

Single-Cell and Single-Molecule Analysis of Gene Expression Regulation

et al. 2016

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Recent advancements in single-cell and single-molecule imaging technologies have resolved biological processes in time and space that are fundamental to understanding the regulation of gene expression. Observations of single-molecule events in their cellular context have revealed highly dynamic aspects of transcriptional and post-transcriptional control in eukaryotic cells. This approach can relate transcription with mRNA abundance and lifetimes. Another key aspect of single-cell analysis is the cell-to-cell variability among populations of cells. Definition of heterogeneity has revealed stochastic processes, determined characteristics of under-represented cell types or transitional states, and integrated cellular behaviors in the context of multicellular organisms. In this review, we discuss novel aspects of gene expression of eukaryotic cells and multicellular organisms revealed by the latest advances in single-cell and single-molecule imaging technology.

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“…In our own laboratory, we are exploiting and further improving Diatrack specifically for tracking dim and highly dynamic mRNA particles during nuclear export – a demanding application for automated tracking technology (see Figure 3E). 106,107 …”

Section: Survey Of Applicationsmentioning

confidence: 99%

Diatrack particle tracking software: Review of applications and performance evaluation

Vallotton

Oijen

Whitchurch

et al. 2017

Traffic

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Object tracking is an instrumental tool supporting studies of cellular trafficking. There are three challenges in object tracking: the identification of targets; the precise determination of their position and boundaries, and the assembly of correct trajectories. This last challenge is particularly relevant when dealing with densely populated images with low signal-to-noise ratios - conditions that are often encountered in applications such as organelle tracking, virus particle tracking, or single-molecule imaging. We have developed a set of methods that can handle a wide variety of signal complexities. They are compiled into a free software package called Diatrack. Here we review its main features and utility in a range of applications, providing a survey of the dynamic imaging field together with recommendations for effective use. The performance of our framework is shown to compare favorably to a wide selection of custom-developed algorithms, whether in terms of localization precision, processing speed, or correctness of tracks.

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In vivo single-particle imaging of nuclear mRNA export in budding yeast demonstrates an essential role for Mex67p

Abstract: Single-particle imaging in budding yeast demonstrates that mRNP export is fast (∼200 ms) and that mRNPs are retained at NPCs and undergo retrograde transport in a mex67-5 mutant, proving an essential role for Mex67p in directional mRNP transport.

Cited by 67 publications

References 61 publications

Structure and Function of the Nuclear Pore Complex Cytoplasmic mRNA Export Platform

Structure and Function of the Nuclear Pore Complex Cytoplasmic mRNA Export Platform

Single-Cell and Single-Molecule Analysis of Gene Expression Regulation

Diatrack particle tracking software: Review of applications and performance evaluation

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