BR-bodies provide selectively permeable condensates that stimulate mRNA decay and prevent release of decay intermediates

Al-Husini, Nadra; Tomares, Dylan T.; Pfaffenberger, Zechariah; Muthunayake, Nisansala S.; Samad, Mohammad Abdus; Zuo, Tiancheng; Bitar, Obaidah; Aretakis, James R.; Bharmal, Mohammed Husain M; Gega, Alisa; Biteen, Julie S.; Childers, W. Seth; Schrader, Jared M.

doi:10.1101/690628

Cited by 21 publications

(55 citation statements)

References 86 publications

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“…For a more in-depth review of BR-body function and phylogenetic distribution, see Muthunayake et al (4). (21) In vitro (18) In vitro (18) In vitro (18) In vitro (22) In vitro (22) In vitro (22) In vitro (47) In vitro (23) In vivo In vitro spherical (23,24,26) In situ icosahedral (95)(96)(97)(98) In situ (25,29) In vitro (23) In vivo (91) In vitro (26) In vitro (23,24,26) ND…”

Section: Current Evidence Demonstrates That Llps May Mediate Subcellumentioning

confidence: 99%

“…In vitro (17,18) In vivo (17) In vivo (31) In vitro (17) proteobacteria; these foci do not form under mRNA depletion (17). Moreover, RNase E CTD foci coalesce into larger spherical structures in Escherichia coli.…”

Section: Current Evidence Demonstrates That Llps May Mediate Subcellumentioning

confidence: 99%

“…Moreover, RNase E CTD foci coalesce into larger spherical structures in Escherichia coli. Also, the ability of BR-bodies to recruit certain RNAs and to exclude the ribosome and nucleoid support their selective permeability (18). This specific recognition may be a result of surface tension controlled by specific mRNA sequences or by mRNA concentration (34).…”

Section: Current Evidence Demonstrates That Llps May Mediate Subcellumentioning

confidence: 99%

“…The liquid-like behavior of condensates has been tested by fluorescence recovery after photobleaching (FRAP) and by observing droplet fusion events (17,21,24,26,28). Finally, (18,21,26,30) and measured the diffusive properties of their molecular components (23,26,31,32). As the methods improve and rigorous controls are implemented, single-molecule methods are becoming increasingly critical for understanding bacterial subcellular organization due to the very small size of the relevant features in these microscopic organisms.…”

Section: Introductionmentioning

confidence: 99%

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The emergence of phase separation as an organizing principle in bacteria

Azaldegui

Vecchiarelli

Biteen

2020

Preprint

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Recent investigations in bacteria suggest that membraneless organelles play a crucial role in the subcellular organization of bacterial cells. However, the biochemical functions and assembly mechanisms of these compartments have not yet been completely characterized. This Review assesses the current methodologies used in the study of membraneless organelles in bacteria, highlights the limitations in determining the phase of complexes in cells that are typically an order of magnitude smaller than a eukaryotic cell, and identifies gaps in our current knowledge about the functional role of membraneless organelles in bacteria. Liquid-liquid phase separation (LLPS) is one proposed mechanism for membraneless organelle assembly. Overall, we outline the framework to evaluate LLPS in vivo in bacteria, we describe the bacterial systems with proposed LLPS activity, and we comment on the general role LLPS plays in bacteria and how it may regulate cellular function. Lastly, we provide an outlook for super-resolution microscopy and single-molecule tracking as tools to assess condensates in bacteria.Statement of SignificanceThough membraneless organelles appear to play a crucial role in the subcellular organization and regulation of bacterial cells, the biochemical functions and assembly mechanisms of these compartments have not yet been completely characterized. Furthermore, liquid-liquid phase separation (LLPS) is one proposed mechanism for membraneless organelle assembly, but it is difficult to determine subcellular phases in tiny bacterial cells. Thus, we outline the framework to evaluate LLPS in vivo in bacteria and we describe the bacterial systems with proposed LLPS activity in the context of these criteria.

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Section: Current Evidence Demonstrates That Llps May Mediate Subcellumentioning

confidence: 99%

Section: Current Evidence Demonstrates That Llps May Mediate Subcellumentioning

confidence: 99%

Section: Current Evidence Demonstrates That Llps May Mediate Subcellumentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The emergence of phase separation as an organizing principle in bacteria

Azaldegui

Vecchiarelli

Biteen

2020

Preprint

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“…We proceeded to characterize the properties of these protein condensates and the dynamics of the encapsulated supercharged GFP. Literature on condensate formation in bacteria remains sparse, but recent reports have shed light on endogenous RNP-bodies (known as BR-bodies) formed in C. crescentus from a protein, RNase E, which contains an unstructured C-terminal domain responsible for intracellular phase separation (6,7). In addition to BRbodies, numerous studies have reported both artificially engineered and common endogenous bacterial compartments (2,11,21,22).…”

Section: Condensed Phase Is Distinct From Inclusion Bodies and Is Dynmentioning

confidence: 99%

Formation of biomolecular condensates in bacteria by tuning protein electrostatics

Yeong

Werth

Brown

et al. 2020

Preprint

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Biomolecular condensates provide a strategy for cellular organization without a physical membrane barrier while allowing for dynamic, responsive organization of the cell. To date, very few biomolecular condensates have been identified in prokaryotes, presenting an obstacle to engineering these compartments in bacteria. As a novel strategy for bacterial compartmentalization, protein supercharging and complex coacervation were employed to engineer liquid-like condensates in E. coli. A simple model for the phase separation of supercharged proteins was developed and used to predict intracellular condensate formation. Herein, we demonstrate that GFP-dense condensates formed by expressing GFP variants of sufficient charge in cells are dynamic and enrich specific nucleic acid and protein components. This study provides a fundamental characterization of intracellular phase separation in E. coli driven by protein supercharging and highlights future utility in designing functional synthetic membraneless organelles.

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Phase‐separated bacterial ribonucleoprotein bodies organize mRNA decay

et al. 2020

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In bacteria, mRNA decay is controlled by megadalton scale macromolecular assemblies called, "RNA degradosomes," composed of nucleases and other RNA decay associated proteins. Recent advances in bacterial cell biology have shown that RNA degradosomes can assemble into phase-separated structures, termed bacterial ribonucleoprotein bodies (BR-bodies), with many analogous properties to eukaryotic processing bodies and stress granules. This review will highlight the functional role that BR-bodies play in the mRNA decay process through its organization into a membraneless organelle in the bacterial cytoplasm. This review will also highlight the phylogenetic distribution of BRbodies across bacterial species, which suggests that these phase-separated structures are broadly distributed across bacteria, and in evolutionarily related mitochondria and chloroplasts.

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BR-bodies provide selectively permeable condensates that stimulate mRNA decay and prevent release of decay intermediates

Cited by 21 publications

References 86 publications

The emergence of phase separation as an organizing principle in bacteria

The emergence of phase separation as an organizing principle in bacteria

Formation of biomolecular condensates in bacteria by tuning protein electrostatics

Phase‐separated bacterial ribonucleoprotein bodies organize mRNA decay

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