ATG9A regulates dissociation of recycling endosomes from microtubules leading to formation of influenza A virus liquid condensates DOI Creative Commons
Sílvia Vale-Costa, Temitope Akhigbe Etibor, Daniela Brás

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Dec. 22, 2022

ABSTRACT It is now established that many viruses threaten public health establish condensates via phase transitions to complete their lifecycles, and knowledge on such processes may offer new strategies for antiviral therapy. In the case of influenza A virus (IAV), liquid known as viral inclusions, concentrate 8 distinct ribonucleoproteins (vRNPs) form IAV genome are viewed sites dedicated assembly 8-partite genomic complex. Despite not being delimited by host membranes, inclusions accumulate membranes inside a result vRNP binding recycling endocytic marker Rab11a, driver biogenesis these structures. We lack molecular understanding how Rab11a-recycling endosomes condensate specifically near endoplasmic reticulum (ER) exit upon infection. show here interact with ER fuse, divide slide. uncover that, contrary previous indications, reported reduction in activity regulated process rather than competition cellular resources involving novel role factor ATG9A. infection, ATG9A mediates removal carrying vRNPs from microtubules. observe usage microtubules rescued when depleted, which prevents condensation Rab11a ER. The failure produce accumulates cytosol, reduces release infectious virions. propose supports dynamics facilitating formation. This work advances our epidemic pandemic genomes formed. also reveals plasticity pathway undergo response disclosing roles beyond its classical involvement autophagy.

Language: Английский

Defining basic rules for hardening influenza A virus liquid condensates DOI Creative Commons
Temitope Akhigbe Etibor, Sílvia Vale-Costa, Sindhuja Sridharan

et al.

eLife, Journal Year: 2023, Volume and Issue: 12

Published: April 4, 2023

In biological systems, liquid and solid-like biomolecular condensates may contain the same molecules but their behaviour, including movement, elasticity, viscosity, is different on account of distinct physicochemical properties. As such, it known that phase transitions affect function material properties can be tuned by several factors temperature, concentration, valency. It is, however, unclear if some are more efficient than others at regulating behaviour. Viral infections good systems to address this question as they form de novo part replication programmes. Here, we used influenza A virus (IAV) cytosolic condensates, AKA viral inclusions, provide a proof concept condensate hardening via changes in valency its components altering concentration or temperature cell. Liquid IAV inclusions hardened targeting vRNP (viral ribonucleoprotein) interactions NP (nucleoprotein) oligomerising molecule, nucleozin, both vitro vivo without affecting host proteome abundance nor solubility. This study starting point for understanding how pharmacologically modulate offer opportunities alternative antiviral strategies.

Language: Английский

Citations

22
Fluorescent protein tags affect the condensation properties of a phase-separating viral protein DOI
Russell J. R. Barkley, Jack C. Crowley, Andrew J. Brodrick

et al.

Molecular Biology of the Cell, Journal Year: 2024, Volume and Issue: 35(7)

Published: May 29, 2024

Fluorescent protein (FP) tags are extensively used to visualize and characterize the properties of biomolecular condensates despite a lack investigation into effects these on phase separation. Here, we characterized dynamic µNS, viral hypothesized undergo separation main component mammalian orthoreovirus factories. Our interest in sequence determinants nucleation process µNS led us compare size density formed by FP::µNS untagged protein. We found an FP-dependent increase droplet density, which suggests that FP can promote condensation. To further assess effect formation, fused mutants show could variably induce otherwise noncondensing proteins. By comparing fluorescent constructs with identified mNeonGreen as least artifactual tag minimally perturbed These results some more suitable for visualizing characterizing minimal experimental artifacts.

Language: Английский

Citations

8
Phosphorylation regulates viral biomolecular condensates to promote infectious progeny production DOI Creative Commons
Nicholas Grams, Matthew Charman, Edwin Halko

et al.

The EMBO Journal, Journal Year: 2024, Volume and Issue: 43(2), P. 277 - 303

Published: Jan. 2, 2024

Abstract Biomolecular condensates (BMCs) play important roles in diverse biological processes. Many viruses form BMCs which have been implicated various functions critical for the productive infection of host cells. The adenovirus L1-52/55 kilodalton protein (52K) was recently shown to viral that coordinate genome packaging and capsid assembly. Although packaging, we do not know how are regulated during infection. Here show phosphorylation serine residues 28 75 within N-terminal intrinsically disordered region 52K modulates vitro cells, promoting liquid-like properties. Furthermore, demonstrate promotes production infectious progeny particles. Collectively, our findings provide insights into condensate properties maintained a state conducive their function production. In addition, implications antiviral strategies aimed at targeting regulation limit multiplication.

Language: Английский

Citations

5
RNA and condensates: Disease implications and therapeutic opportunities DOI Creative Commons

Tina W. Han,

Bede Portz,

Richard A. Young

et al.

Cell chemical biology, Journal Year: 2024, Volume and Issue: 31(9), P. 1593 - 1609

Published: Sept. 1, 2024

Language: Английский

Citations

5
Localising enzymes to biomolecular condensates increases their accumulation and benefits engineered metabolic pathway performance in Nicotiana benthamiana DOI Creative Commons
Anya Lindström Battle, A. John Barrett, Mark D. Fricker

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 29, 2025

Summary Establishment of N. benthamiana as a robust biofactory is complicated by issues such product toxicity and proteolytic degradation target proteins / introduced enzymes. Here we investigate whether biomolecular condensates can be used to address these problems. We engineered in leaves using transient expression synthetic modular scaffolds. The vivo properties the that resulted were consistent with them being liquid-like bodies thermodynamic features typical multicomponent phase-separating systems. show recruitment enzymes led several fold yield increases one- three-step metabolic pathways (citramalate biosynthesis poly-3-hydroxybutyrate (PHB) biosynthesis, respectively). This enhanced could for reasons including improved enzyme kinetics, metabolite channelling or avoidance cytotoxicity retention pathway within condensate, which was demonstrated PHB. However, also observed several-fold increase amount accumulated when they targeted condensates. suggests more stable localised condensate than freely diffusing cytosol. hypothesise this stability likely main driver increased production. Our findings provide foundation leveraging plant engineering advance versatile industrial applications.

Language: Английский

Citations

0
Fluorescence Loss After Photoactivation (FLAPh): A Pulse-Chase Cellular Assay for Understanding Kinetics and Dynamics of Viral Inclusions DOI
Temitope Akhigbe Etibor, Tiago Paixão, Maria João Amorim

et al.

Methods in molecular biology, Journal Year: 2025, Volume and Issue: unknown, P. 125 - 140

Published: Jan. 1, 2025

Language: Английский

Citations

0
O-GlcNAcylation reduces proteome solubility and regulates the formation of biomolecular condensates in human cells DOI Creative Commons
Senhan Xu, Kejun Yin, Xing Xu

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: April 30, 2025

O-GlcNAcylation plays critical roles in the regulation of protein functions and cellular activities, including interactions with other macromolecules. While formation biomolecular condensates (or biocondensates) regulated by a few individual proteins has been reported, systematic investigation on biocondensate remains to be explored. Here we systematically study regulating solubility its impacts RNA-protein using mass spectrometry-based chemoproteomics. Unexpectedly, observe system-wide decrease modified O-GlcNAcylation, glycoproteins involved focal adhesion actin binding exhibiting most significant decrease. Furthermore, sites located disordered regions fewer acidic aromatic residues nearby are related greater drop solubility. Additionally, discover that specific group events promotes dissociation under heat stress, while some enhance during recovery phase. Using site mutagenesis, inhibition O-GlcNAc transferase, fluorescence microscopy, validate regulates biocondensates for YTHDF3 NUFIP2. This work advances our understanding condensates.

Language: Английский

Citations

0
Viral amyloids: New opportunities for antiviral therapeutic strategies DOI Creative Commons
Frank Gondelaud, Pierre‐Yves Lozach, Sonia Longhi

et al.

Current Opinion in Structural Biology, Journal Year: 2023, Volume and Issue: 83, P. 102706 - 102706

Published: Sept. 30, 2023

Language: Английский

Citations

10
ATG9A regulates the dissociation of recycling endosomes from microtubules to form liquid influenza A virus inclusions DOI Creative Commons
Sílvia Vale-Costa, Temitope Akhigbe Etibor, Daniela Brás

et al.

PLoS Biology, Journal Year: 2023, Volume and Issue: 21(11), P. e3002290 - e3002290

Published: Nov. 20, 2023

It is now established that many viruses threaten public health establish condensates via phase transitions to complete their lifecycles, and knowledge on such processes may offer new strategies for antiviral therapy. In the case of influenza A virus (IAV), liquid known as viral inclusions, concentrate 8 distinct ribonucleoproteins (vRNPs) form IAV genome are viewed sites dedicated assembly 8-partite genomic complex. Despite not being delimited by host membranes, inclusions accumulate membranes inside a result vRNP binding recycling endocytic marker Rab11a, driver biogenesis these structures. We lack molecular understanding how Rab11a-recycling endosomes condensate specifically near endoplasmic reticulum (ER) exit upon infection. show here interact with ER fuse, divide, slide. uncover that, contrary previous indications, reported reduction in activity regulated process rather than competition cellular resources involving novel role factor ATG9A. infection, ATG9A mediates removal carrying vRNPs from microtubules. observe usage microtubules rescued when depleted, which prevents condensation Rab11a ER. The failure produce accumulates cytosol reduces release infectious virions. propose supports dynamics facilitating formation. This work advances our epidemic pandemic genomes formed. also reveals plasticity undergo response disclosing roles beyond its classical involvement autophagy.

Language: Английский

Citations

10
Biomolecular condensates with liquid properties formed during viral infections DOI Creative Commons
Damien Glon,

Benjamin Léonardon,

Ariane Guillemot

et al.

Microbes and Infection, Journal Year: 2024, Volume and Issue: 26(8), P. 105402 - 105402

Published: Aug. 8, 2024

During a viral infection, several membraneless compartments with liquid properties are formed. They can be of origin concentrating proteins and nucleic acids, harboring essential stages the cycle, or cellular containing components involved in innate immunity. This is paradigm shift our understanding replication interaction between viruses

Language: Английский

Citations

3