scholarly journals Translational regulation of protrusion-localized RNAs involves silencing and clustering after transport

2019 ◽  
Author(s):  
Konstadinos Moissoglu ◽  
Kyota Yasuda ◽  
Tianhong Wang ◽  
George Chrisafis ◽  
Stavroula Mili
Keyword(s):  
Gene Expression ◽  
Translational Regulation ◽  
Cellular Responses ◽  
Distinct Mode ◽  
Control Of Gene Expression ◽  
Spatial Control ◽  
Polarized Cells ◽  
Regulate Protein ◽  
In Transit ◽  
Human And Mouse

ABSTRACTLocalization of RNAs to various subcellular destinations is a widely used mechanism that regulates a large proportion of transcripts in polarized cells. In many cases, such localized transcripts mediate spatial control of gene expression by being translationally silent while in transit and locally activated at their destination. Here, we investigate the translation of RNAs localized at dynamic cellular protrusions of human and mouse, migrating, mesenchymal cells. In contrast to the model described above, we find that protrusion-localized RNAs are not locally activated solely at protrusions, but can be translated with similar efficiency in both internal and peripheral locations. Interestingly, protrusion-localized RNAs are translated at extending protrusions, they become translationally silenced in retracting protrusions and this silencing is accompanied by coalescence of single RNAs into larger heterogeneous RNA clusters. This work describes a distinct mode of translational regulation of localized RNAs, which we propose is used to regulate protein activities during dynamic cellular responses.

scholarly journals Translational regulation of protrusion-localized RNAs involves silencing and clustering after transport

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Konstadinos Moissoglu ◽  
Kyota Yasuda ◽  
Tianhong Wang ◽  
George Chrisafis ◽  
Stavroula Mili
Keyword(s):  
Gene Expression ◽  
Translational Regulation ◽  
Cellular Responses ◽  
Distinct Mode ◽  
Control Of Gene Expression ◽  
Spatial Control ◽  
Polarized Cells ◽  
Regulate Protein ◽  
In Transit ◽  
Human And Mouse

Localization of RNAs to various subcellular destinations is a widely used mechanism that regulates a large proportion of transcripts in polarized cells. In many cases, such localized transcripts mediate spatial control of gene expression by being translationally silent while in transit and locally activated at their destination. Here, we investigate the translation of RNAs localized at dynamic cellular protrusions of human and mouse, migrating, mesenchymal cells. In contrast to the model described above, we find that protrusion-localized RNAs are not locally activated solely at protrusions, but can be translated with similar efficiency in both internal and peripheral locations. Interestingly, protrusion-localized RNAs are translated at extending protrusions, they become translationally silenced in retracting protrusions and this silencing is accompanied by coalescence of single RNAs into larger heterogeneous RNA clusters. This work describes a distinct mode of translational regulation of localized RNAs, which we propose is used to regulate protein activities during dynamic cellular responses.

scholarly journals A versatile genetic tool for post-translational control of gene expression in Drosophila melanogaster

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Sachin Sethi ◽  
Jing W Wang
Keyword(s):  
Gene Expression ◽  
Side Effects ◽  
Translational Control ◽  
High Efficiency ◽  
Developmental Stages ◽  
Dynamic Range ◽  
In Vivo Studies ◽  
Control Of Gene Expression ◽  
Regulate Protein

Several techniques have been developed to manipulate gene expression temporally in intact neural circuits. However, the applicability of current tools developed for in vivo studies in Drosophila is limited by their incompatibility with existing GAL4 lines and side effects on physiology and behavior. To circumvent these limitations, we adopted a strategy to reversibly regulate protein degradation with a small molecule by using a destabilizing domain (DD). We show that this system is effective across different tissues and developmental stages. We further show that this system can be used to control in vivo gene expression levels with low background, large dynamic range, and in a reversible manner without detectable side effects on the lifespan or behavior of the animal. Additionally, we engineered tools for chemically controlling gene expression (GAL80-DD) and recombination (FLP-DD). We demonstrate the applicability of this technology in manipulating neuronal activity and for high-efficiency sparse labeling of neuronal populations.

scholarly journals A versatile genetic tool for post-translational control of gene expression in Drosophila melanogaster

2017 ◽  
Author(s):  
Sachin Sethi ◽  
Jing W. Wang
Keyword(s):  
Gene Expression ◽  
Side Effects ◽  
Translational Control ◽  
High Efficiency ◽  
Developmental Stages ◽  
Dynamic Range ◽  
Control Of Gene Expression ◽  
Neuronal Populations ◽  
Regulate Protein

AbstractSeveral techniques have been developed to manipulate gene expression temporally in intact neural circuits. However, the applicability of current tools developed for in vivo studies in Drosophila is limited by their incompatibility with existing GAL4 lines and side effects on physiology and behavior. To circumvent these limitations, we adopted a strategy to reversibly regulate protein degradation with a small molecule by using a destabilizing domain (DD). We show that this system is effective across different tissues and developmental stages. We further show that this system can be used to control in vivo gene expression levels with low background, large dynamic range, and in a reversible manner without detectable side effects on the lifespan or behavior of the animal. Additionally, we engineered tools for chemically controlling gene expression (GAL80-DD) and recombination (FLP-DD). We demonstrate the applicability of this technology in manipulating neuronal activity and for high-efficiency sparse labeling of neuronal populations.

scholarly journals Spatial Control of Gene Expression by miR319-Regulated TCP Transcription Factors in Leaf Development

2017 ◽  
Vol 176 (2) ◽  
pp. 1694-1708 ◽  
Author(s):  
Edgardo G. Bresso ◽  
Uciel Chorostecki ◽  
Ramiro E. Rodriguez ◽  
Javier F. Palatnik ◽  
Carla Schommer
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Leaf Development ◽  
Control Of Gene Expression ◽  
Spatial Control

scholarly journals Spatial control of gene expression within a scaffold by localized inducer release

Biomaterials ◽  
2011 ◽  
Vol 32 (11) ◽  
pp. 3062-3071 ◽  
Author(s):  
Priya R. Baraniak ◽  
Devin M. Nelson ◽  
Cory E. Leeson ◽  
Anand K. Katakam ◽  
Jennifer L. Friz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Control Of Gene Expression ◽  
Spatial Control

scholarly journals CreLite: An Optogenetically Controlled Cre/loxP System Using Red Light

2019 ◽  
Author(s):  
Shuo-Ting Yen ◽  
Kenneth A. Trimmer ◽  
Nader Aboul ◽  
Rachel D. Mullen ◽  
James C. Culver ◽  
...  
Keyword(s):  
Gene Expression ◽  
Light Exposure ◽  
Red Light ◽  
Regulatory Elements ◽  
Cell Labeling ◽  
P System ◽  
Zebrafish Embryos ◽  
Control Of Gene Expression ◽  
Spatial Control ◽  
Temporal And Spatial

ABSTRACTPrecise manipulation of gene expression with temporal and spatial control is essential for functional studies and the determination of cell lineage relationships in complex biological systems. The Cre-loxP system is commonly used for gene manipulation at desired times and places. However, specificity is dependent on the availability of tissue- or cell-specific regulatory elements used in combination with Cre or CreER (tamoxifen-inducible). Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos. Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6. In addition, PhyB-PIF6 binding requires phycocyanobilin (PCB), providing an additional layer of control. Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity. Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter (ubi:zebrabow) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium. We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development. CreLite provides a novel optogenetic tool for precise temporal and spatial control of gene expression in zebrafish embryos that may also be useful in cell culture, ex vivo organ culture, and other animal models for developmental biology studies.

Translational control in the C. elegans hermaphrodite germ line

Genome ◽  
2010 ◽  
Vol 53 (2) ◽  
pp. 83-102 ◽  
Author(s):  
Hilary Racher ◽  
Dave Hansen
Keyword(s):  
Gene Expression ◽  
Cell Fate ◽  
Translational Control ◽  
Translational Regulation ◽  
Developmental Stages ◽  
Germ Line ◽  
Control Of Gene Expression ◽  
Cell Fate Decisions ◽  
C Elegans ◽  
Mrna Targets

The formation of a fully developed gamete from an undifferentiated germ cell requires progression through numerous developmental stages and cell fate decisions. The precise timing and level of gene expression guides cells through these stages. Translational regulation is highly utilized in the germ line of many species, including Caenorhabditis elegans , to regulate gene expression and ensure the proper formation of gametes. In this review, we discuss some of the developmental stages and cell fate decisions involved in the formation of functional gametes in the C. elegans germ line in which translational control has been implicated. These stages include the mitosis versus meiosis decision, the sperm/oocyte decision, and gamete maturation. We also discuss some of the techniques used to identify mRNA targets; the identification of these targets is necessary to clearly understand the role each RNA-binding protein plays in these decisions. Relatively few mRNA targets have been identified, thus providing a major focus for future research. Finally, we propose some reasons why translational control may be utilized so heavily in the germ line. Given that many species have this substantial reliance on translational regulation for the control of gene expression in the germ line, an understanding of translational regulation in the C. elegans germ line is likely to increase our understanding of gamete formation in general.

scholarly journals A conserved choreography of mRNAs at centrosomes reveals a localization mechanism involving active polysome transport

2020 ◽  
Author(s):  
Adham Safieddine ◽  
Emeline Coleno ◽  
Abdel-Meneem Traboulsi ◽  
Oh Sung Kwon ◽  
Frederic Lionneton ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
High Throughput ◽  
Local Translation ◽  
Protein Coding ◽  
Control Of Gene Expression ◽  
Spatial Control ◽  
Centrosomal Protein ◽  
Protein Coding Genes ◽  
Drug Treatments

AbstractLocal translation allows for a spatial control of gene expression. Here, we used high-throughput smFISH to screen centrosomal protein-coding genes, and we describe 8 human mRNAs accumulating at centrosomes. These mRNAs localize at different stages during cell cycle with a remarkable choreography, indicating a finely regulated translational program at centrosomes. Interestingly, drug treatments and reporter analyses revealed a common translation-dependent localization mechanism requiring the nascent protein. Using ASPM and NUMA1 as models, single mRNA and polysome imaging revealed active movements of endogenous polysomes towards the centrosome at the onset of mitosis, when these mRNAs start localizing. ASPM polysomes associate with microtubules and localize by either motor-driven transport or microtubule pulling. Remarkably, the Drosophila orthologs of the human centrosomal mRNAs also localize to centrosomes and also require translation. These data identify a conserved family of centrosomal mRNAs that localize by active polysomes transport mediated by nascent proteins.

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