Single-base m6A epitranscriptomics reveals novel HIV-1 host interaction targets in primary CD4+ T cells

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

Published: Jan. 2, 2025

Abstract N 6 -methyladenosine (m A) is the most prevalent cellular mRNA modification and plays a critical role in regulating RNA stability, localization, gene expression. m A vital modulating expression of viral genes during HIV-1 infection. infection increases levels many cell types, which facilitates replication infectivity target cells. However, function primary CD4 + T cells remains unclear. Here, we demonstrate that Jurkat promotes interaction between writer complex subunits methyltransferase-like 3 14 (METTL3/METTL14). Using single-base A-specific sequencing, identified several differentially A-modified mRNAs, including perilipin ( PLIN3 ), Interestingly, increased level by enhancing its but protein was decreased. Knocking down reduced production enhanced virion infectivity. In contrast, cells, were unaffected infection, knocking out did not impact or These results indicate interplay cell-type specific only observed Overall, our highlight importance HIV-1-infected suggest significance as regulatory mechanism Author Summary common chemical on helps control important for this study, found two proteins, METTL3 METTL14, are responsible adding modifications to RNA. mRNAs with altered one called . stabilized levels, When knocked it decreased made particles more infectious. line, affect knockout alter infectivity, suggesting effect Our findings show host like unique infected

Language: Английский

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An HIV-1 Reference Epitranscriptome

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

Published: Jan. 31, 2025

ABSTRACT Post-transcriptional chemical modifications to RNA, or the epitranscriptome, play important roles in RNA metabolism, gene regulation, and human disease, including viral pathogenesis. Modifications genome transcripts of immunodeficiency virus 1 (HIV-1) have been reported, methylation adenosine (m 6 A) cytosine 5 C), acetylation cytosine, pseudouridylation (psi), conversion inosine, their effects on host biology investigated. However, diverse experimental approaches used, making clear correlations across studies difficult assess. To address this need, we propose establishment a reference HIV-1 epitranscriptome. We sequenced model NL4-3 from infected Jurkat CD4+ T cells using latest nanopore chemistry, custom preparation methods, commercial base-calling algorithms. This resulted reproducible sense preliminary antisense epitranscriptome where m A, C, psi, ands inosine could be identified by multiplexed base-calling. Multiplexed miscalled due sequence neighboring modification contexts, which demonstrate can corrected with synthetic fragments. validate A sites small molecule inhibitor methyltransferase-like 3 (METTL3), STM2457. conclude that do not change substantially under combination antiretroviral therapy (cART) treatment primary cells. Samples patients living HIV reveal conservation certain modifications, such as A. Our approach data offer straightforward benchmark adopted help advance rigor, reproducibility, uniformity future epitranscriptomics studies.

Language: Английский

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Dynamic interplay between RNA N6-methyladenosine modification and porcine reproductive and respiratory syndrome virus infection

Veterinary Research, Journal Year: 2025, Volume and Issue: 56(1)

Published: March 22, 2025

Abstract N 6 -methyladenosine (m A) has attracted significant attention for its role in regulating the complex interaction between viruses and host cells. Porcine reproductive respiratory syndrome virus (PRRSV) is a pathogen affecting swine health worldwide. Here, we first identified seven m A-enriched peaks PRRSV genomic RNA by A immunoprecipitation sequencing A-seq). Moreover, functional analyses revealed positive correlation modification level replication. Treatment with universal methylation inhibitor 3-deazaadenosine (3-DAA) effectively suppressed replication dose-dependent manner. Furthermore, A-seq was also used to determine landscape of transcriptome PAMs infected pandemic or highly pathogenic strains. Among 4677 transcripts exhibiting altered levels, MAPK14 gene p38/MAPK signalling pathway emerged as preliminary targets A-mediated epigenetic regulation during infection. These findings provide new insights into mechanisms underlying infection may facilitate development anti-PRRSV therapeutics.

Language: Английский

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RNA-Seq analysis reveals the long noncoding RNAs associated with immunity in wild Myotis myotis bats

BMC Genomics, Journal Year: 2025, Volume and Issue: 26(1)

Published: April 5, 2025

Bats possess a uniquely adapted immune system that enables them to live with viral infections without the expected maladies. The molecular basis and regulation of bats' response is still not fully understood. Long non-coding RNAs (lncRNAs) represent an emerging class molecules critical regulatory roles in multiple biological processes, including immunity. We hypothesise lncRNA-based bats may enable limit disease pathogens. developed lncRNA prediction pipeline annotate long transcriptome across bat tissues at population level. Characterisation our dataset based on 100 blood transcriptomes from wild Myotis myotis revealed lower more tissue-specific expression compared coding genes, reduced GC content shorter length distributions, consistent profiles observed other species. Using WGCNA network analyses gene ontology, we identified two mRNA-lncRNA co-expression modules associated distinct response: one linked T-cell activation vial inflammation. From these immune-related lncRNAs, selected four candidates high translational potential for regulating These include newly lncRNA, BatLnc1, antiviral functions; M. ortholog TUG1, implicated viral-host interactions; well-known lncRNAs MALAT1 NEAT1, recognised their inflammatory regulation. conducted first ab initio non-model species, wild-caught myotis. Our analysis significant variation status among subset individuals, potentially due pathogenic conditions. variations, most likely bats. This initial exploration lays groundwork future experimental validations functions, offering promising insights into role

Language: Английский

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Inhibition of ALKBH5 demethylase of m6A pathway potentiates HIV-1 reactivation from latency

Virology Journal, Journal Year: 2025, Volume and Issue: 22(1)

Published: April 28, 2025

Language: Английский

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