HIVToolbox, an Integrated Web Application for Investigating HIV
(A) Phosphorylation of IN by CK2. Reactions containing IN, 32P-γ-ATP and CK2 were serially diluted 1∶3 starting from 50 U/mL (right lane). The extent of phosphorylation was visualized by SDS-PAGE and autoradiography. (B) Phosphorylation of IN domains by CK2. Reactions as in A and contained purified GST-IN or GST-IN domains as indicated in the diagram. IN(S283A) is a point mutation of one of the putative CKII phosphorylation sites. (C) His-tagged point mutants of putative CKII sites within IN were phosphorylated by CK2 as in (A). A control reaction lacking IN showed no phosphorylation (not shown). Reduced phosphorylation of S283A mutant is indicated (*). Samples were normalized for protein concentration.