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Fig. 2 | Virology Journal

Fig. 2

From: Noncovalent SUMO-interaction motifs in HIV integrase play important roles in SUMOylation, cofactor binding, and virus replication

Fig. 2

IN SIMs are required for the IN-SUMO2/3 interaction. a Schematics of various plasmid constructs encoding GFP-INwt/mut, HA-SUMO2, HA-SUMO3 and HA-ubc9. b 293 T cells were cotransfected with GFP-INwt/IN3VI and HA-SUMO2/3, the IN/SUMO interaction was analyzed by co-IP assay. Upper Panel: The HA-SUMO3/2 bound GFP-IN; Middle pane: The expression of GFP-INwt/3VI; Lower panel: The expression of HA-SUMO3/2 in 2% of total cell lysates. c SIM2 and SIM3 of IN mediate IN-SUMO3 interaction. Two hundred and ninety-three T cells were cotransfected with GFP-INwt/mut and HA-SUMO3 as indicated. Upper panel: The HA-SUMO3 bound GFP-INwt/mut; Middle and lower panels: The GFP-INwt/mut and HA-SUMO3 present in the 2% of the total cell lysates (middle panel on the left). d The relative ratio of pulled down GFP-INmut to GFP-INwt was calculated through densitometric analysis of the autoradiograms on c. The binding affinity of GFP-IN wt and HA-SUMO3 was arbitrarily set as 100%. The results are representative of two independent experiments

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