DNA‐dependent phase separation by human SSB2 (NABP1/OBFC2A) protein points to adaptations to eukaryotic genome repair processes DOI Creative Commons
Zoltán J. Kovács, Gábor M. Harami, János Pálinkás

et al.

Protein Science, Journal Year: 2024, Volume and Issue: 33(4)

Published: March 21, 2024

Single-stranded DNA binding proteins (SSBs) are ubiquitous across all domains of life and play essential roles via stabilizing protecting single-stranded (ss) as well organizing multiprotein complexes during replication, recombination, repair. Two mammalian SSB paralogs (hSSB1 hSSB2 in humans) were recently identified shown to be involved various genome maintenance processes. Following our recent discovery the liquid-liquid phase separation (LLPS) propensity Escherichia coli (Ec) SSB, here we show that also forms LLPS condensates under physiologically relevant ionic conditions. Similar seen for EcSSB, demonstrate contribution hSSB2's C-terminal intrinsically disordered region (IDR) condensate formation, selective enrichment metabolic condensates. However, contrast EcSSB-driven is inhibited by ssDNA binding, requires nucleic acid especially facilitated ssDNA. Our results reveal an evolutionarily conserved role SSB-mediated spatiotemporal organization complexes. At same time, differential features EcSSB point functional adaptations prokaryotic versus eukaryotic contexts.

Language: Английский

Revolutionizing viral disease treatment: Phase separation and lysosome/exosome targeting as new areas and new paradigms for antiviral drug research DOI
Shaoqing Du, Xueping Hu, Xinyong Liu

et al.

Drug Discovery Today, Journal Year: 2024, Volume and Issue: 29(3), P. 103888 - 103888

Published: Jan. 21, 2024

Language: Английский

Citations

5
Plant viruses and biomolecular condensates: novel perspectives in virus replication strategies DOI Open Access
Jared P. May

New Phytologist, Journal Year: 2024, Volume and Issue: 243(5), P. 1636 - 1638

Published: April 24, 2024

This article is a Commentary on Lin & Nagy (2024), 243 : 1917–1935.

Language: Английский

Citations

5
Biomolecular phase separation in stress granule assembly and virus infection DOI Creative Commons
Yi Liu,

Zhiying Yao,

Guiwei Lian

et al.

Acta Biochimica et Biophysica Sinica, Journal Year: 2023, Volume and Issue: 55(7), P. 1099 - 1118

Published: June 14, 2023

Liquid-liquid phase separation (LLPS) has emerged as a crucial mechanism for cellular compartmentalization. One prominent example of this is the stress granule. Found in various types cells, granule biomolecular condensate formed through separation. It comprises numerous RNA and RNA-binding proteins. Over past decades, substantial knowledge been gained about composition dynamics granules. SGs can regulate signaling pathways have associated with human diseases, such neurodegenerative cancer, infectious diseases. The threat viral infections continues to loom over society. Both DNA viruses depend on host cells replication. Intriguingly, many stages life cycle are closely tied metabolism cells. field condensates rapidly advanced recent times. In context, we aim summarize research granules their link infections. Notably, triggered by behave differently from canonical sodium arsenite (SA) heat shock. Studying context could offer valuable platform replication processes anti-viral responses. A deeper understanding these biological pave way innovative interventions treatments They potentially bridge gap between basic interactions hosts.

Language: Английский

Citations

13
G-quadruplexes control hepatitis B virus replication by promoting cccDNA transcription and phase separation in hepatocytes DOI Creative Commons
Guillaume Giraud, Mélanie Rodà, Pélagie Huchon

et al.

Nucleic Acids Research, Journal Year: 2023, Volume and Issue: 52(5), P. 2290 - 2305

Published: Dec. 19, 2023

Abstract Phase separation regulates fundamental processes in gene expression and is mediated by the local concentration of proteins nucleic acids, as well acid secondary structures such G-quadruplexes (G4s). These play roles both host viral replication due to their peculiar localisation regulatory sequences. Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) an episomal minichromosome whose persistence at basis chronic infection. Identifying mechanisms controlling its transcriptional activity indispensable develop new therapeutic strategies against hepatitis B. The aim this study was determine whether G4s are formed cccDNA regulate replication. Combining biochemistry functional studies, we demonstrate that indeed contains ten structures. Furthermore, mutations disrupting two located enhancer I HBV region altered transcription Finally, showed for first time undergoes phase a G4-dependent manner promote infected hepatocytes. Altogether, our data give insight regulation might pave way identification novel targets destabilize or silence cccDNA.

Language: Английский

Citations

12
Abnormal phase separation of biomacromolecules in human diseases DOI
Songhao Zhang, Gaofeng Pei, Boya Li

et al.

Acta Biochimica et Biophysica Sinica, Journal Year: 2023, Volume and Issue: 55(7), P. 1133 - 1152

Published: July 1, 2023

Membrane-less organelles (MLOs) formed through liquid-liquid phase separation (LLPS) are associated with numerous important biological functions, but the abnormal will also dysregulate physiological processes. Emerging evidence points to importance of LLPS in human health and diseases. Nevertheless, despite recent advancements, our knowledge molecular relationship between diseases is frequently incomplete. In this review, we outline current understanding about how aberrant affects developmental disorders, tandem repeat cancers viral infection. We examine disease mechanisms driven by condensates, highlight potential treatment approaches. This study seeks expand providing a valuable new paradigm for as well further translate regarding into therapeutic discoveries.

Language: Английский

Citations

11
Buffer choice and pH strongly influence phase separation of SARS-CoV-2 nucleocapsid with RNA DOI
Nina C. Kathe, Mihajlo Novakovic, Frédéric H.‐T. Allain

et al.

Molecular Biology of the Cell, Journal Year: 2024, Volume and Issue: 35(5)

Published: April 3, 2024

The SARS-CoV-2 nucleocapsid (N) protein is crucial for virus replication and genome packaging. N forms biomolecular condensates both in vitro vivo a process known as liquid–liquid phase separation (LLPS), but the exact factors regulating LLPS of are not fully understood. Here, we show that pH buffer choice have profound impact on protein. degree highly dependent solution, which correlated with histidine protonation Specifically, demonstrate H356 essential phosphate buffer. Moreover, electrostatic interactions molecules specific amino acid residues able to alter net charge protein, thus influencing its ability undergo presence RNA. Overall, these findings reveal even subtle changes or surface caused by system can strongly influence behavior, point main driving forces separation. Further, our emphasize importance experimental parameters when studying biomolecules, especially context viral infections where intracellular milieu undergoes drastic normally decreases.

Language: Английский

Citations

4
Attenuation of viral replication foci in nuclei by 1,6 Hexanediol implicates phase separation in the assembly of baculoviral replication factories DOI
Alexander D. Finoshin, О. И. Кравчук, K. I. Adameyko

et al.

Journal of Virological Methods, Journal Year: 2025, Volume and Issue: 335, P. 115147 - 115147

Published: March 6, 2025

Language: Английский

Citations

0
Influenza A virus RNA localisation and the interceding trafficking pathways of the host cell DOI Creative Commons
Stefano Bonazza, David G. Courtney

PLoS Pathogens, Journal Year: 2025, Volume and Issue: 21(4), P. e1013090 - e1013090

Published: April 23, 2025

Viruses have evolved to efficiently navigate host cells deliver, express, and replicate their genetic material. Understanding the mechanisms underlying viral RNA localisation is paramount designing new antivirals. In this review, we discuss Influenza A Virus (IAV) as a model system highlight some of ways in which viruses can hijack endomembrane systems, well nuclear transporters, achieve correct transcripts. IAV exemplifies nuclear-replicating virus with complex highly regulated trafficking within cells. The subverts various vesicular transport systems altering normal cellular functions. begins during entry; after clathrin-mediated endocytosis, genome (vRNPs) released into cytosol fusion endosomal membrane, it subsequently imported nucleus via importin system. There, vRNPs engage most major subnuclear structures exploit chromatin, transcription machinery splicing apparatus efficient mRNA synthesis export. Subsequently, newly synthesised are rapidly exported from contact host’s recycling endosome network for plasma membrane. We critical remodelling entire system, particularly Rab11 endoplasmic reticulum. Lastly, replicated genomes come together bundles be inserted budding virions, current models being proposed evidence behind them. Despite advances understanding these processes, several knowledge gaps remain, regarding specific export unspliced transcripts, segment bundling.

Language: Английский

Citations

0
Liquid–liquid phase separation of LARP7 restrains HIV-1 replication DOI Creative Commons
Zhuoxin Li, Xiya Fang, Bing Zhao

et al.

EMBO Reports, Journal Year: 2025, Volume and Issue: unknown

Published: March 20, 2025

Abstract HIV-1 initiates replication by its transactivator Tat, hijacking the positive transcription elongation factor b (P-TEFb) in host cell. Most P-TEFb is maintained an inactive state 7SK snRNP until it brought to initiation complex cellular or viral transactivators that accelerate and facilitate production of full-length transcripts. Here, we report infection triggers liquid-liquid phase separation LARP7, a central component snRNP. Tat incorporated into HIV-1-induced LARP7 condensates after infection. Conserved lysine residues intrinsically disordered region are essential for both inhibition Tat-mediated transcription. These findings identify mechanism wherein sequestered within condensates, restraining

Language: Английский

Citations

0
Pseudorabies virus IE180 protein hijacks G3BPs into the nucleus to inhibit stress granule formation DOI Creative Commons

Ruihan Zhao,

Zhenbang Zhu, Wenqiang Wang

et al.

Journal of Virology, Journal Year: 2025, Volume and Issue: unknown

Published: March 27, 2025

ABSTRACT Pseudorabies virus (PRV) is a porcine alphaherpesvirus that can infect different animal species and cause pruritus lethal encephalitis. Stress granules (SGs) are membrane-free cytoplasmic structures formed by liquid-liquid phase separation of G3BP proteins during cell translation inhibition, which generally plays an antiviral role in various viral infections. In this study, we found infection with PRV strains inhibits the formation SGs host cells. We IE180, only immediate early protein PRV, has distinct inhibitory effect on SG colocalizes SG-nucleating (G3BP1/2) nucleus infection. Co-immunoprecipitation assays demonstrated interaction between IE180 G3BP1/G3BP2, appears to depend Herpesvirus ICP4-like N-terminal (ICP4L-N) domain both NTF2L RBD domains G3BP1. Since G3BPs mainly function cytoplasm induce formation, constructed several truncations lacking nuclear localization sequence alter subcellular cytoplasm. Mutant was expressed still suppressed induced arsenite or poly(I:C), but its ability inhibit weakened. Importantly, knockout facilitated replication H1299 cells, while exogenous expression wild-type cells replication. summary, our study indicates suppresses hijacks into benefit IMPORTANCE Herpesviruses, including pseudorabies (PRV), have evolved strategies compromise immune responses. one targets viruses overcome order increase The related herpes simplex 1 (HSV-1) promote replication, underlying mechanisms remain unknown. confirmed formation. Interestingly, interacts these prevent line G3BP1/2 significantly compromised reported mechanism be also utilized HSV-1 Therefore, elucidates novel alphaherpesviruses provides new perspective inquire escape other alphaherpesviruses.

Language: Английский

Citations

0