Accession	GSE250106
Status	2024/5/20
Title	Epitranscriptomic cytidine methylation of the Hepatitis B viral RNA encapsidation signal ensure the reverse transcription of viral RNA [PA-m5C-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.
Samples	GSM ID Sample info Characteristics Description GSM7974002 HuH-7, mock transfected, PAm5Cseq cell line:HuH-7; cell type:hepatocyte; genotype:WT; treatment:untreated - GSM7974003 HuH-7, HBV transfected, PAm5Cseq cell line:HuH-7; cell type:hepatocyte; genotype:WT; treatment:+HBV - GSM7974004 CRISPR control HuH-7, HBV transfected, PAm5Cseq cell line:HuH-7; cell type:hepatocyte; genotype:non-targeted Cas9 stable line; treatment:+HBV - GSM7974005 NSUN2-KO HuH-7, HBV transfected, PAm5Cseq cell line:HuH-7; cell type:hepatocyte; genotype:NSUN2 knockout; treatment:+HBV -
Platform	GPL16791  GPL18573  : Illumina HiSeq 2500 (Homo sapiens);Illumina NextSeq 500 (Homo sapiens)
Literature	38830096