Development and Characterization of an Oncolytic Human Adenovirus-Based Vector Co-Expressing the Adenovirus Death Protein and p14 Fusion-Associated Small Transmembrane Fusogenic Protein DOI Open Access

Kathy L. Poulin,

Ryan G. Clarkin, Joshua Del Papa

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(22), P. 12451 - 12451

Published: Nov. 20, 2024

Human adenovirus (HAdV)-based oncolytic vectors, which are designed to preferentially replicate in and kill cancer cells, have shown modest efficacy human clinical trials part due poor viral distribution throughout the tumor mass. Previously, we showed that expression of p14 fusion-associated small transmembrane (FAST) fusogenic protein could enhance HAdV reduce growth rate a xenograft mouse model cancer. We now explore whether co-expression death (ADP) with FAST synergize further vector efficacy. ADP is naturally encoded within early region 3 (E3) HAdV, frequently removed from HAdV-based functions cell lysis progeny release. evaluated variety approaches achieve optimal two proteins, most efficient method being insertion an cassette E3 deletion, consisting coding sequences for separated by self-cleaving peptide derived porcine teschovirus-1 (P2A). However, quantities produced this were reduced approximately 10-fold compared similar vector-expressing only wildtype respectively. Compared our original alone, P2A construct cell-cell fusion, spread, cell-killing activity A549 adenocarcinoma cells culture. These studies show can be used express different transgenes armed vector, but also highlight challenges maintaining adequate transgene when modifying design.

Language: Английский

Restrictor slows early transcription elongation to render RNA polymerase II susceptible to termination at non-coding RNA loci DOI Creative Commons
Claudia A. Mimoso, Hanneke Vlaming,

Nathalie P. de Wagenaar

et al.

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

Published: Jan. 10, 2025

The eukaryotic genome is broadly transcribed by RNA polymerase II (RNAPII) to produce protein-coding messenger RNAs (mRNAs) and a repertoire of non-coding (ncRNAs). Whereas RNAPII very processive during mRNA transcription, it terminates rapidly synthesis many ncRNAs, particularly those that arise opportunistically from accessible chromatin at gene promoters or enhancers. divergent fates versus ncRNA species raise questions about how associated machineries discriminate functional spurious transcription. Restrictor complex, comprised the binding protein ZC3H4 RNAPII-interacting WDR82, has been implicated in restraining expression ncRNAs. However, determinants targeting mechanism transcription suppression remain unclear. Here, we investigate using unbiased sequence screens, rapid degradation followed nascent sequencing. We find promiscuously suppresses early elongation RNAPII, but this activity blocked most mRNAs presence 5' splice site. Consequently, critical determinant directionality prevents transcriptional interference. Finally, our data indicate rather than directly terminating acts reducing rate elongation, rendering susceptible termination other machineries.

Language: Английский

Citations

0
Epitranscriptomics in the Glioma Context: A Brief Overview DOI Open Access
Pablo Santamarina‐Ojeda, Agustín F. Fernández, Mario F. Fraga

et al.

Cancers, Journal Year: 2025, Volume and Issue: 17(4), P. 578 - 578

Published: Feb. 8, 2025

Epitranscriptomics, the study of chemical modifications in RNA, has emerged as a crucial field cellular regulation, adding another layer to established landscape DNA- and histone-based epigenetics. A wide range RNA modifications, including N6-methyladenosine, pseudouridine, inosine, have been identified across nearly all species, influencing essential processes such transcription, splicing, stability, translation. In context brain tumors, particularly gliomas, specific epitranscriptomic signatures reported play role tumorigenesis. Despite growing evidence, biological implications various remain poorly understood. This review offers an examination main interplay between modified unmodified molecules, how they could contribute glioma-like phenotypes, therapeutic impact targeting these mechanisms.

Language: Английский

Citations

0
Dynamics and evolutionary conservation of B complex protein recruitment during spliceosome activation DOI Creative Commons
Xingyang Fu, Aaron A. Hoskins

Nucleic Acids Research, Journal Year: 2025, Volume and Issue: 53(4)

Published: Feb. 8, 2025

Abstract Spliceosome assembly and catalytic activation involve dozens of protein snRNA binding unbinding events. The B-complex specific proteins (Prp38, Snu23, Spp381) have critical roles in stabilizing the spliceosome during rearrangements essential for activation. While these are conserved, different mechanisms been proposed their recruitment to spliceosomes. To visualize directly, we used Colocalization Single Molecule Spectroscopy (CoSMoS) study dynamics Prp38, Spp381 splicing real time. These can bind release from spliceosomes simultaneously likely associated with one another. We designate minimally containing as a potential B complex (BCP) subcomplex spliceosome. Under conditions, BCP associate pre-mRNA after tri-snRNP binding. predominantly occurs U4 snRNP dissociation NineTeen Complex (NTC) association. low concentrations ATP, preassociate resulting simultaneous pre-mRNA. Together, our results reveal that is conserved between Saccharomyces cerevisiae humans. when ATP limiting may result formation unproductive complexes could be regulate splicing.

Language: Английский

Citations

0
Understanding GEMIN5 Interactions: From Structural and Functional Insights to Selective Translation DOI Creative Commons
Encarnación Martı́nez-Salas, Salvador Abellan, Rosario Francisco‐Velilla

et al.

Wiley Interdisciplinary Reviews - RNA, Journal Year: 2025, Volume and Issue: 16(2)

Published: March 1, 2025

ABSTRACT GEMIN5 is a predominantly cytoplasmic protein, initially identified as member of the survival motor neurons (SMN) complex. In addition, this abundant protein modulates diverse aspects RNA‐dependent processes, executing its functions through formation multi‐component complexes. The modular organization structural domains present in enables to perform various interaction with distinct partners. responsible for recognition small nuclear (sn)RNAs N‐terminal region, and therefore snRNP assembly. Beyond role spliceosome assembly, regulates translation either RNAs or proteins. central robust dimerization domain acts hub protein–protein interaction, while non‐canonical RNA‐binding site located towards C‐terminus. Interestingly, partitioning mRNAs into polysomes, likely due capacity ability bind native ribosomes. Understanding functional has brought an increasing interest last years important implications human disease. Patients carrying biallelic variants suffer from neurodevelopmental delay, hypotonia, cerebellar ataxia. This review discusses recent relevant works aimed at understanding molecular mechanisms activity gene expression, also challenges discover new functions.

Language: Английский

Citations

0
TAF2 condensation in nuclear speckles links basal transcription factor TFIID to RNA splicing factors DOI
Tanja Bhuiyan,

Niccolò Arecco,

Paulina Karen Mendoza Sanchez

et al.

Cell Reports, Journal Year: 2025, Volume and Issue: 44(5), P. 115616 - 115616

Published: April 26, 2025

Language: Английский

Citations

0
Chromatin-associated lncRNA-splicing factor condensates regulate hypoxia responsive RNA processing of genes pre-positioned near nuclear speckles DOI Open Access
You Jin Song, Min Kyung Shinn, Sushant Bangru

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 31, 2024

Abstract Hypoxia-induced alternative splicing (AS) regulates tumor progression and metastasis. Little is known about how such AS controlled whether higher-order genome nuclear domain (ND) organizations dictate these processes. We observe that hypoxia-responsive alternatively spliced genes position near speckle (NS), the ND enhances efficiency. NS-resident MALAT1 long noncoding RNA, induced in response to hypoxia, AS. achieves this by organizing SR-family of factor, SRSF1, NS regulating binding SRSF1 pre-mRNAs. Mechanistically, recruitment elongating RNA polymerase II (pol II) promoting formation phase-separated condensates which are preferentially recognized pol II. During spatially organized establishing a threshold concentration NSs, potentially forming condensates, critical for II-mediated

Language: Английский

Citations

1
Circadian PERIOD proteins sculpt the mammalian alternative splicing landscape DOI Open Access
Lies Chikhaoui,

K Mamgain,

Masahide Seki

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 23, 2024

Abstract Mammalian circadian oscillators are driven by a transcription-translation feedback loop where CLOCK:BMAL1 activity is repressed the PER:CRY complex. While transcriptional regulation PER well established, role of in co- and post- processes remains unclear. Here, we used Nanopore long-read direct RNA sequencing (dRNAseq) quantitative mass spectrometry (qMS) to uncover critical function PERs alternative splicing (AS) liver. Our expanded transcriptome revealed significant changes rhythmic expression annotated transcripts, novel isoforms known genes, previously unannotated with widespread perturbations Per1 -/- ;Per2 (PerKO) livers. Rhythmic AS events were restricted distinct subset entropy - metric complexity displayed oscillations only limited number pathways, primarily those associated glucose homeostasis cellular responses insulin. In PerKO livers, however, detected increased isoform altered across broad range pathways linked cell growth, morphogenesis, ER-associated degradation (ERAD), insulin response histone methylation. Biochemical analyses qMS data indicate that these not due mis-expression factors, but rather stem from nuclear abundance chromatin retention few Serine-Arginine-rich factors (SRSFs). particular, SRSF3 acts proximal core-clock defining both period amplitude rhythms. findings highlight for proteins shaping liver proteome integrating transcription complex dynamic landscape.

Language: Английский

Citations

1
Development and Characterization of an Oncolytic Human Adenovirus-Based Vector Co-Expressing the Adenovirus Death Protein and p14 Fusion-Associated Small Transmembrane Fusogenic Protein DOI Open Access

Kathy L. Poulin,

Ryan G. Clarkin, Joshua Del Papa

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(22), P. 12451 - 12451

Published: Nov. 20, 2024

Human adenovirus (HAdV)-based oncolytic vectors, which are designed to preferentially replicate in and kill cancer cells, have shown modest efficacy human clinical trials part due poor viral distribution throughout the tumor mass. Previously, we showed that expression of p14 fusion-associated small transmembrane (FAST) fusogenic protein could enhance HAdV reduce growth rate a xenograft mouse model cancer. We now explore whether co-expression death (ADP) with FAST synergize further vector efficacy. ADP is naturally encoded within early region 3 (E3) HAdV, frequently removed from HAdV-based functions cell lysis progeny release. evaluated variety approaches achieve optimal two proteins, most efficient method being insertion an cassette E3 deletion, consisting coding sequences for separated by self-cleaving peptide derived porcine teschovirus-1 (P2A). However, quantities produced this were reduced approximately 10-fold compared similar vector-expressing only wildtype respectively. Compared our original alone, P2A construct cell-cell fusion, spread, cell-killing activity A549 adenocarcinoma cells culture. These studies show can be used express different transgenes armed vector, but also highlight challenges maintaining adequate transgene when modifying design.

Language: Английский

Citations

0