{"doi":"10.1101/2020.11.04.368712","title":"<i>N</i> <sup>6</sup> -methyladenosine modification of HIV-1 RNA evades RIG-I-mediated sensing to suppresses type-I interferon induction in monocytic cells","abstract":"Abstract N 6 -methyladenosine (m 6 A) is a prevalent RNA modification that plays a key role in regulating eukaryotic cellular mRNA functions. RNA m 6 A modification is regulated by two groups of cellular proteins, writers and erasers that add or remove m 6 A, respectively. HIV-1 RNA contains m 6 A modifications that modulate viral infection and gene expression in cells. However, it remains unclear whether m 6 A modifications of HIV-1 RNA modulate innate immune responses in cells or HIV-1-infected individuals. Here we show that m 6 A modification of HIV-1 RNA suppresses the expression of antiviral cytokine type-I interferon (IFN-I) in human monocytic cells. Transfection of differentiated monocytic cells with HIV-1 RNA fragments containing a single m 6 A-modification significantly reduced IFN-I mRNA expression relative to their unmodified RNA counterparts. We generated HIV-1 with altered RNA m 6 A levels by manipulating the expression of the m 6 A erasers or pharmacological inhibition of m 6 A addition in virus-producing cells. RNA transfection and viral infection of differentiated monocytic cells demonstrated that HIV-1 RNA with decreased m 6 A levels enhanced IFN-I expression, whereas HIV-1 RNA with increased m 6 A modifications had opposite effects. Our mechanistic studies revealed that m 6 A of HIV-1 RNA escaped the RIG-I-mediated RNA sensing and activation of the transcription factors IRF3 and IRF7 that drive IFN-I gene expression. Moreover, RNA of peripheral blood mononuclear cells from HIV-1 viremic patients showed increased m 6 A levels that correlated with increased IFN-I mRNA expression compared to levels from HIV-1-suppressed patients on antiretroviral therapy. Together, our results suggest that RNA m 6 A modifications regulate viral replication and antiviral innate immune responses in HIV-1-infected individuals. Author Summary HIV-1 is known as a weak inducer of antiviral cytokines including IFN-I, but it is unclear how HIV-1 evades innate immunity. Different types of RNA modifications including m 6 A within the HIV-1 genome modulate viral replication; however, the role of m 6 A modifications of HIV-1 RNA in regulating innate immune responses remains elusive. In this study, we found that HIV-1 RNA modified with m 6 A suppresses the expression of IFN-I in differentiated monocytic cells by avoiding innate immune detection of viral RNA mediated by RIG-I, an RNA sensor in host cells. We also observed significantly increased RNA m 6 A modifications of peripheral blood mononuclear cells from HIV-1 viremic patients compared to virally suppressed patients on combined antiretroviral therapy, suggesting a functional link between m 6 A modifications and antiretroviral treatment. Investigating the functions of m 6 A modifications of HIV-1 RNA in regulating innate immune sensing and IFN-I induction in monocytic cells can help understand the mechanisms of HIV-1 persistence.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":131421,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9515,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":585415,"name":"Sameer Kumar","orcid":"0000-0001-8697-7370","position":1,"is_corresponding":false},{"id":585416,"name":"Nagaraja Tirumuru","orcid":"0000-0001-9042-5210","position":2,"is_corresponding":false},{"id":497564,"name":"Jennifer L. Welch","orcid":"0000-0003-2725-9875","position":3,"is_corresponding":false},{"id":239950,"name":"Lulu Hu","orcid":"0000-0001-7246-1736","position":4,"is_corresponding":false},{"id":585417,"name":"Chuan He","orcid":"0000-0003-0965-7984","position":5,"is_corresponding":false},{"id":332249,"name":"Jack T. Stapleton","orcid":"0000-0002-2302-9055","position":6,"is_corresponding":false},{"id":576943,"name":"Li Wu","orcid":"0000-0002-5468-2487","position":7,"is_corresponding":false},{"id":121773,"name":"Shuliang Chen","orcid":"0000-0002-7175-7604","position":0,"is_corresponding":true}],"reference_count":56,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:16:03.875886Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}