Mitotic Kinases Aurora-A, Plk1, and Cdk1 Interact with Elk-1 Transcription Factor through the N-Terminal Domain

Figure 2

Interaction of mitotic kinases with Elk-1 protein. (a–c) Domain of interaction on Elk-1 protein: (a) a schematic diagram of GST-Elk-1 deletion mutants in pGEX-2T vector used in this study; (b) GST pulldown analysis of Elk-1 and mitotic kinase interaction; GST alone (pGEX) or GST-Elk-1 deletions were expressed in BL21 pLysS strain and semipurified using glutathione-sepharose beads, followed by incubation with U87 glioblastoma cell lysates; pulldown samples were analyzed with Western blot using primary antibodies specific for Aur-A, Aur-B, Cdk1, Plk1, ERK, and SRF; GST-Elk-1 deletion inputs were analyzed with GST antibody (lower panel); (c) putative binding motifs (blue line for Plk1, green line for Cdk1, and red line for Aurora-A or Aurora-B) on Elk-1 protein sequence for the mitotic kinases and their predicted phosphorylation sites (blue font for Plk1, green font for Cdk1, and red font for Aurora-A or Aurora-B) within amino acids 93-205 (red box).

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