Long-Term Fluorescence Recovery After Photobleaching (FRAP)

Saitô, Tunezô; Matsunaga, Daiki; Deguchi, Shinji

doi:10.1007/978-1-0716-2851-5_21

Cited by 2 publications

(3 citation statements)

References 21 publications

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“…A small region of the droplets is irradiated with light until the fluorophore is destroyed and the region is bleached. Since photobleaching is irreversible, the observed signal of fluorescence recovery is attributable to the exchange of bleached fluorophores with the unbleached ones present in the surrounding area . Passive transport processes, such as Brownian motions, create a net transfer of molecules out and into the region of interest until equilibrium.…”

Section: Methods To Study Rna Crowdingmentioning

confidence: 99%

“…Since photobleaching is irreversible, the observed signal of fluorescence recovery is attributable to the exchange of bleached fluorophores with the unbleached ones present in the surrounding area. 233 Passive transport processes, such as Brownian motions, create a net transfer of molecules out and into the region of interest until equilibrium. The measure of how rapidly (or how slowly) the initial fluorescent is recovered, in full or partially, is correlated with the fluidity of the droplet.…”

Section: Micropipette Aspiration Assay Maintaining a Different Range ...mentioning

confidence: 99%

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RNA: The Unsuspected Conductor in the Orchestra of Macromolecular Crowding

Zacco,

Broglia,

Kurihara

et al. 2024

Chem. Rev.

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This comprehensive Review delves into the chemical principles governing RNA-mediated crowding events, commonly referred to as granules or biological condensates. We explore the pivotal role played by RNA sequence, structure, and chemical modifications in these processes, uncovering their correlation with crowding phenomena under physiological conditions. Additionally, we investigate instances where crowding deviates from its intended function, leading to pathological consequences. By deepening our understanding of the delicate balance that governs molecular crowding driven by RNA and its implications for cellular homeostasis, we aim to shed light on this intriguing area of research. Our exploration extends to the methodologies employed to decipher the composition and structural intricacies of RNA granules, offering a comprehensive overview of the techniques used to characterize them, including relevant computational approaches. Through two detailed examples highlighting the significance of noncoding RNAs, NEAT1 and XIST, in the formation of phase-separated assemblies and their influence on the cellular landscape, we emphasize their crucial role in cellular organization and function. By elucidating the chemical underpinnings of RNAmediated molecular crowding, investigating the role of modifications, structures, and composition of RNA granules, and exploring both physiological and aberrant phase separation phenomena, this Review provides a multifaceted understanding of the intriguing world of RNA-mediated biological condensates.

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Section: Methods To Study Rna Crowdingmentioning

confidence: 99%

Section: Micropipette Aspiration Assay Maintaining a Different Range ...mentioning

confidence: 99%

RNA: The Unsuspected Conductor in the Orchestra of Macromolecular Crowding

Zacco,

Broglia,

Kurihara

et al. 2024

Chem. Rev.

View full text Add to dashboard Cite

show abstract

“…This gradually leads to a third, stationary phase, where further association and dissociation of monomers at both ends of the filament are in equilibrium with each other (treadmilling phase) [ 63 ]. In cells, therefore, individual filaments, or at least networks and bundles, display continuous turnover, although turnover rates in different subcellular actin structures are highly variable [ 64 , 65 ]. Importantly, G-actin has an intrinsic polarity that is translated into the filament, leading consequently to two filament ends with differential growth features.…”

Section: Correlating Cytochalasan Chemodiversity With the Activity Sp...mentioning

confidence: 99%

Cytochalasans and Their Impact on Actin Filament Remodeling

et al. 2023

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The eukaryotic actin cytoskeleton comprises the protein itself in its monomeric and filamentous forms, G- and F-actin, as well as multiple interaction partners (actin-binding proteins, ABPs). This gives rise to a temporally and spatially controlled, dynamic network, eliciting a plethora of motility-associated processes. To interfere with the complex inter- and intracellular interactions the actin cytoskeleton confers, small molecular inhibitors have been used, foremost of all to study the relevance of actin filaments and their turnover for various cellular processes. The most prominent inhibitors act by, e.g., sequestering monomers or by interfering with the polymerization of new filaments and the elongation of existing filaments. Among these inhibitors used as tool compounds are the cytochalasans, fungal secondary metabolites known for decades and exploited for their F-actin polymerization inhibitory capabilities. In spite of their application as tool compounds for decades, comprehensive data are lacking that explain (i) how the structural deviances of the more than 400 cytochalasans described to date influence their bioactivity mechanistically and (ii) how the intricate network of ABPs reacts (or adapts) to cytochalasan binding. This review thus aims to summarize the information available concerning the structural features of cytochalasans and their influence on the described activities on cell morphology and actin cytoskeleton organization in eukaryotic cells.

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Long-Term Fluorescence Recovery After Photobleaching (FRAP)

Cited by 2 publications

References 21 publications

RNA: The Unsuspected Conductor in the Orchestra of Macromolecular Crowding

RNA: The Unsuspected Conductor in the Orchestra of Macromolecular Crowding

Cytochalasans and Their Impact on Actin Filament Remodeling

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