title: N(6)-methyladenosine of HIV-1 RNA regulates viral infection and HIV-1 Gag protein expression.
abstract: The internal N(6)-methyladenosine (m(6)A) methylation of eukaryotic nuclear RNA controls post-transcriptional gene expression, which is regulated by methyltransferases (writers), demethylases (erasers), and m(6)A-binding proteins (readers) in cells. The YTH domain family proteins (YTHDF1-3) bind to m(6)A-modified cellular RNAs and affect RNA metabolism and processing. Here, we show that YTHDF1-3 proteins recognize m(6)A-modified HIV-1 RNA and inhibit HIV-1 infection in cell lines and primary CD4(+) T-cells. We further mapped the YTHDF1-3 binding sites in HIV-1 RNA from infected cells. We found that the overexpression of YTHDF proteins in cells inhibited HIV-1 infection mainly by decreasing HIV-1 reverse transcription, while knockdown of YTHDF1-3 in cells had the opposite effects. Moreover, silencing the m(6)A writers decreased HIV-1 Gag protein expression in virus-producing cells, while silencing the m(6)A erasers increased Gag expression. Our findings suggest an important role of m(6)A modification of HIV-1 RNA in viral infection and HIV-1 protein synthesis.
authors.affiliation: Center for Retrovirus Research, The Ohio State University, Columbus, United States.|Department of Veterinary Biosciences, The Ohio State University, Columbus, United States.
authors.forename: Nagaraja
authors.initials: N
authors.lastname: Tirumuru
authors.affiliation: Department of Chemistry, The University of Chicago, Chicago, United States.|Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, United States.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, United States.|Howard Hughes Medical Institute, The University of Chicago, Chicago, United States.
authors.forename: Boxuan Simen
authors.initials: BS
authors.lastname: Zhao
authors.affiliation: Center for Retrovirus Research, The Ohio State University, Columbus, United States.|Department of Veterinary Biosciences, The Ohio State University, Columbus, United States.
authors.forename: Wuxun
authors.initials: W
authors.lastname: Lu
authors.affiliation: Department of Chemistry, The University of Chicago, Chicago, United States.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, United States.|Howard Hughes Medical Institute, The University of Chicago, Chicago, United States.|Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, United States.
authors.forename: Zhike
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Department of Chemistry, The University of Chicago, Chicago, United States.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, United States.|Howard Hughes Medical Institute, The University of Chicago, Chicago, United States.|Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, United States.
authors.forename: Chuan
authors.initials: C
authors.lastname: He
authors.affiliation: Center for Retrovirus Research, The Ohio State University, Columbus, United States.|Department of Veterinary Biosciences, The Ohio State University, Columbus, United States.|Department of Microbial Infection and Immunity, The Ohio State University, Columbus, United States.|Comprehensive Cancer Center, The Ohio State University, Columbus, United States.
authors.forename: Li
authors.identifier: 0000-0002-5468-2487
authors.initials: L
authors.lastname: Wu
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism; D002478:Cells, Cultured; D015967:Gene Expression Regulation, Viral; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development*; D054884:Host-Pathogen Interactions; D006801:Humans; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis*
chemical_list: D012367:RNA, Viral; D016601:RNA-Binding Proteins; C000600306:YTHDF1 protein, human; C585900:YTHDF2 protein, human; C000625021:YTHDF3 protein, human; D054301:gag Gene Products, Human Immunodeficiency Virus; C010223:N-methyladenosine; D000241:Adenosine
keywords: HIV-1; N6-methyladenosine; RNA methylation; YTHDF proteins; human; infectious disease; microbiology; replication; virus