Fig. 1: Mutation of R18G and K25A profoundly reduces infection but differentially impacts capsid formation. | Nature Communications

Fig. 1: Mutation of R18G and K25A profoundly reduces infection but differentially impacts capsid formation.

From: HIV-1 adapts to lost IP6 coordination through second-site mutations that restore conical capsid assembly

Fig. 1

A Cross-section of a CA hexamer showing two IP6 molecules bound within the pore by the charged rings at R18 and K25 (based on 6R6Q). Side panel shows a close-up view. B Titration of WT and mutant HIV-1 VSV-G-pseudotyped virus on HEK293T cells with infectivity quantified as the proportion of infected cells (area of monolayer). Error bars depict mean ± s.e.m. from at least three independent experiments (N = 3). C Cryo-ET analysis of the indicated HIV-1 mutants. Tilt series were collected and reconstructions performed to assess capsid morphology. A total of 51 WT, 38 K25A and 43 R18G particles were analyzed and classified into the indicated categories. Example sliced tomograms belonging to each category are shown together with the data for each virion. Scale bars, 100 nm. D In vitro assembly of 900 µM CA (WT, K25A and R18G) in 6 mM IP6, measuring absorbance of reaction over time at 350 nm. Negative stain EM images of assembly reactions are shown with 4x zoomed in sections displayed above. Scalebar: 200 nm.

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