HIV protease cleaves the antiviral m6A reader protein YTHDF3 in the viral particle
Fig 5
Cleavage ofYTHDF3 in protease-resistant HIV mutant strains.
(A) Western blot of concentrated virus lysates produced in HEK293TΔYTH3 cells co-transfected with FLAG-YTHDF3 and HIV NL4-3Δenv WT, NL4-3Δenv-MDR1 (Indinavir resistance associated mutations I84V & L90M), NL4-3Δenv-MDR2 (L90M), or control plasmid. Increasing doses of Indinavir or DMSO-only control was added to cells one day after transfection. Membranes were probed with anti-YTHDF3 ab103328 and anti-Gag p24. Data shown is representative of three individual experiments. (B) Quantification of virion incorporated full-length YTHDF3 protein (64 kD, YTHDF364) and the YTH3 fragment (44 kD, YTHDF344) as determined by Western blot (e.g., as shown in Fig 5A). Percent YTHDF364 was calculated relative to the total YTHDF364 and YTHDF344 in each lane. Data shown for two independent experiments are shown. The DMSO-only control is artificially placed at 0.0016 μM for the purpose of graphical representation.