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N 6 -Methyladenosine-binding proteins suppress HIV-1 infectivity and viral production
Journal article   Open access   Peer reviewed

N 6 -Methyladenosine-binding proteins suppress HIV-1 infectivity and viral production

Wuxun Lu, Nagaraja Tirumuru, Corine St Gelais, Pratibha C Koneru, Chang Liu, Mamuka Kvaratskhelia, Chuan He and Li Wu
The Journal of biological chemistry, Vol.293(34), pp.12992-13005
08/24/2018
DOI: 10.1074/jbc.RA118.004215
PMCID: PMC6109920
PMID: 29976753
url
https://doi.org/10.1074/jbc.RA118.004215View
Published (Version of record) Open Access

Abstract

The internal -methyladenosine (m A) modification of cellular mRNA regulates post-transcriptional gene expression. The YTH domain family proteins (YTHDF1-3 or Y1-3) bind to m A-modified cellular mRNAs and modulate their metabolism and processing, thereby affecting cellular protein translation. We previously reported that HIV-1 RNA contains the m A modification and that Y1-3 proteins inhibit HIV-1 infection by decreasing HIV-1 reverse transcription activity. Here, we investigated the mechanisms of Y1-3-mediated inhibition of HIV-1 infection in target cells and the effect of Y1-3 on viral production levels in virus-producing cells. We found that Y1-3 protein overexpression in HIV-1 target cells decreases viral genomic RNA (gRNA) levels and inhibits both early and late reverse transcription. Purified recombinant Y1-3 proteins preferentially bound to the m A-modified 5' leader sequence of gRNA compared with its unmodified RNA counterpart, consistent with the strong binding of Y1-3 proteins to HIV-1 gRNA in infected cells. HIV-1 mutants with two altered m A modification sites in the 5' leader sequence of gRNA exhibited significantly lower infectivity than WT, replication-competent HIV-1, confirming that these sites alter viral infection. HIV-1 produced from cells in which endogenous Y1, Y3, or Y1-3 proteins were knocked down singly or together had increased viral infectivity compared with HIV-1 produced in control cells. Interestingly, we found that Y1-3 proteins and HIV-1 Gag protein formed a complex with RNA in HIV-1-producing cells. Overall, these results indicate that Y1-3 proteins inhibit HIV-1 infection and provide new insights into the mechanisms by which the m A modification of HIV-1 RNA affects viral replication.
HIV-1 - metabolism HIV Infections - virology Humans Virion - metabolism Virus Internalization Gene Products, gag - metabolism HIV-1 - growth & development Adenosine - analogs & derivatives Adenosine - metabolism Protein Binding RNA, Viral - metabolism HeLa Cells Virion - growth & development RNA-Binding Proteins - metabolism HIV Infections - metabolism

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