N6-methyladenosine of HIV-1 RNA regulates viral infection and HIV-1 Gag protein expression
Nagaraja Tirumuru,
Boxuan Simen Zhao,
Wuxun Lu,
Zhike Lu,
Chuan He,
Li Wu
Affiliations
Nagaraja Tirumuru
Center for Retrovirus Research, The Ohio State University, Columbus, United States; Department of Veterinary Biosciences, The Ohio State University, Columbus, United States
Boxuan Simen Zhao
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
Wuxun Lu
Center for Retrovirus Research, The Ohio State University, Columbus, United States; Department of Veterinary Biosciences, The Ohio State University, Columbus, United States
Zhike Lu
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
Chuan He
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
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
The internal N6-methyladenosine (m6A) methylation of eukaryotic nuclear RNA controls post-transcriptional gene expression, which is regulated by methyltransferases (writers), demethylases (erasers), and m6A-binding proteins (readers) in cells. The YTH domain family proteins (YTHDF1–3) bind to m6A-modified cellular RNAs and affect RNA metabolism and processing. Here, we show that YTHDF1–3 proteins recognize m6A-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 m6A writers decreased HIV-1 Gag protein expression in virus-producing cells, while silencing the m6A erasers increased Gag expression. Our findings suggest an important role of m6A modification of HIV-1 RNA in viral infection and HIV-1 protein synthesis.
