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Series GSE250107 Query DataSets for GSE250107
Status Public on May 20, 2024
Title Epitranscriptomic cytidine methylation of the Hepatitis B viral RNA encapsidation signal ensure the reverse transcription of viral RNA [PA-m6A-seq]
Organism Homo sapiens
Experiment type Other
Summary Epitranscriptomic RNA modifications have emerged as important regulators of the fate and function of both cellular and viral RNAs. One prominent modification, the cytidine methylation 5-methylcytidine (m5C), is found on the RNA of HIV-1, where m5C enhances the translation and splicing of HIV-1 RNA. However, whether m5C functionally enhances the RNA of other pathogenic viruses remain elusive. Here, we report that the RNA of hepatitis B virus (HBV) is enriched with a high level of m5C, mediated mainly through the cellular methyltransferase NSUN2. Intrigingly, the most prominent cluster of NSUN2-deposited m5C is found on the epsilon hairpin, an RNA element required for viral RNA encapsidation and reverse transcription. Loss of m5C from HBV RNA due to depletion of NSUN2 resulted in a modest decrease in viral capsid protein (HBc) translation, yet this is accompaneied by a near-complete loss of the reverse transcribed viral DNA. Similarly, mutations introduced to remove the methylated cytidines resulted in a translation decrease and block of reverse transcription. Furthermore, pharmacological disruption of m5C deposition with a nucleoside analogue led to a significant decrease in HBV replication. Thus, our data indicates m5C methylations is a critical enhancer of the epsilon element in HBV reverse transcription. Our study suggests the theraputic potential of targeting the m5C methyltransfer process on the HBV 5’epsilon as an alternative antiviral stratagy.
 
Overall design PA-m6A-seq: Photoactivated UV crosslinking of m56Aantibody to m6A+ RNA, followed by immunoprecipitation and sequencing. RNA from HuH-7 cells, WT or NSUN2 knockout cells, transfected with or without HBV replicon plasmid.
 
Contributor(s) Su P, Tsai K
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Submission date Dec 13, 2023
Last update date May 20, 2024
Contact name Kevin Tsai
E-mail(s) kevtsai@ibms.sinica.edu.tw
Organization name Academia Sinica
Department Institute of Biomedical Sciences
Lab R414
Street address 128 Sec. 2, Academia Rd. Nankang
City Taipei
ZIP/Postal code 115
Country Taiwan
 
Platforms (1)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
Samples (2)
GSM7974006 HuH-7, mock transfected, PAm6Aseq
GSM7974007 HuH-7, HBV transfected, PAm6Aseq
This SubSeries is part of SuperSeries:
GSE250109 Epitranscriptomic cytidine methylation of the Hepatitis B viral RNA encapsidation signal ensure the reverse transcription of viral RNA
Relations
BioProject PRJNA1052036

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Supplementary file Size Download File type/resource
GSE250107_RAW.tar 30.0 Kb (http)(custom) TAR (of BED)
SRA Run SelectorHelp
Raw data are available in SRA

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