Figure 4
From: HIV-1 capsid undergoes coupled binding and isomerization by the nuclear pore protein NUP358
FIV uncouples binding and isomerization by NUP358. (a) ITC trace of NUP358Cyp interaction with FIV CAN. (b) Overlay of 2D 1H-15N ZZ-exchange spectra of FIV CAN in the presence of NUP358Cyp at 3 and 197 ms mixing times. Zoomed-in views of the peaks corresponding to G87 and R89 are also shown. The cross-hairs indicate where exchange peaks would be situated, if cis-trans isomerization took place. (c) Overlay of uniformly 15N-labeled and selective 15N-Arg labeled FIV CAN. Spectra of selective 15N-Arg labeled FIV CAN are also shown in panels on the right in the absence (top) and presence (bottom) of human NUP358Cyp. (d) Upper: Western blots showing depletion of NUP358 or TNPO3 in HeLa cells using antibodies against NUP358, TNPO3 or β-Actin as loading control [6]. Lower: VSV-G pseudotyped GFP-encoding HIV-1 or FIV infection of HeLa cells and HeLa cells expressing scrambled control (shC) or NUP358 or TNPO3 specific shRNA [6, 33] (mean of three independent viral doses, error bars show standard deviation). Titers are plotted as infectious units per ng of reverse transcriptase activity. Data are representative of two independent experiments.