Figure 4.
JIL-1 interactions with the MSL complex are mediated by the kinase domains. (A) Diagram of the 1,207-residue JIL-1 double kinase and the five GST fusion constructs used for pull-down assays. A nearly full-length (GST–JIL-1) and four truncated (GST-NTD, GST-KDI, GST-KDII, and GST-CTD) GST–JIL-1 fusion proteins were generated. The JIL-1 amino acid residues included in each construct are indicated at the right. (B) S2 cell lysate incubated with each of the JIL-1 GST fusion constructs diagrammed in (A) or a GST-only control was pelleted with glutathione-agarose beads and the interacting proteins fractionated by SDS-PAGE, Western blotted, and probed with anti-MSL1 (top panel) or anti-MSL3 (bottom panel) antibody. Unincubated S2 cell lysate was included as a control (lane 1). GST–JIL-1 (lane 3) as well as constructs containing each of the kinase domains, GST-KDI (lane 5) and GST-KDII (lane 6), were able to pull down the MSL complex as indicated by detection with both MSL1 and MSL3 antibody, while no interaction was observed with the GST-only control (lane 2) or the GST-NTD (lane 4) or GST-CTD (lane 7) constructs.