HIV reprograms host m6Am RNA methylome by viral Vpr protein-mediated degradation of PCIF1

Abstract

N⁶,2'-O-dimethyladenosine (m⁶Am) is an abundant RNA modification located adjacent to the 5'-end of the mRNA 7-methylguanosine (m⁷G) cap structure. m⁶A methylation on 2'-O-methylated A at the 5'-ends of mRNAs is catalyzed by the methyltransferase Phosphorylated CTD Interacting Factor 1 (PCIF1). The role of m⁶Am and the function of PCIF1 in regulating host-pathogens interactions are unknown. Here, we investigate the dynamics and reprogramming of the host m⁶Am RNA methylome during HIV infection. We show that HIV infection induces a dramatic decrease in m⁶Am of cellular mRNAs. By using PCIF1 depleted T cells, we identify 2237 m⁶Am genes and 854 are affected by HIV infection. Strikingly, we find that PCIF1 methyltransferase function restricts HIV replication. Further mechanism studies show that HIV viral protein R (Vpr) interacts with PCIF1 and induces PCIF1 ubiquitination and degradation. Among the m⁶Am genes, we find that PCIF1 inhibits HIV infection by enhancing a transcription factor ETS1 (ETS Proto-Oncogene 1, transcription factor) stability that binds HIV promoter to regulate viral transcription. Altogether, our study discovers the role of PCIF1 in HIV-host interactions, identifies m⁶Am modified genes in T cells which are affected by viral infection, and reveals how HIV regulates host RNA epitranscriptomics through PCIF1 degradation.