Nicotinamide adenine dinucleotide (NAD) is a critical component of the cellular metabolism and also serves as an alternative 5' cap on various RNAs. However, the function of the NAD RNA cap is still under investigation. We studied NAD capping of RNAs in HIV-1-infected cells because HIV-1 is responsible for the depletion of the NAD/NADH cellular pool and causing intracellular pellagra. By applying the NAD captureSeq protocol to HIV-1-infected and uninfected cells, we revealed that four snRNAs (e.g., U1) and four snoRNAs lost their NAD cap when infected with HIV-1. Here, we provide evidence that the presence of the NAD cap decreases the stability of the U1/HIV-1 pre-mRNA duplex. Additionally, we demonstrate that reducing the quantity of NAD-capped RNA by overexpressing the NAD RNA decapping enzyme DXO results in an increase in HIV-1 infectivity. This suggests that NAD capping is unfavorable for HIV-1 and plays a role in its infectivity.

1st Faculty of Medicine Charles University Kateřinská 32 121 08 Prague Czechia

Department of Molecular Biology and Genetics Faculty of Science University of South Bohemia Branišovská 1760 37005 České Budějovice Czechia

Division of Biophysics Faculty of Physics University of Warsaw Pasteura 5 02 093 Warsaw Poland

Faculty of Science Department of Cell Biology Charles University Viničná 7 121 08 Prague 2 Czechia

Faculty of Science Department of Physical and Macromolecular Chemistry Charles University Hlavova 8 121 08 Prague 2 Czechia

Institute of Molecular Genetics of the Czech Academy of Sciences Vídeňská 1083 142 20 Prague 4 Czechia

Institute of Organic Chemistry and Biochemistry of the CAS Flemingovo náměstí 2 160 00 Prague 6 Czechia

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The Mysterious World of Non-Canonical Caps - What We Know and Why We Need New Sequencing Techniques

If the 5' cap fits (wear it) - Non-canonical RNA capping