Skip to main content
. 2001 Dec;12(12):3987–3999. doi: 10.1091/mbc.12.12.3987

Figure 5.

Figure 5

CDK6, KSHV-cyclin, and p16 form a ternary complex. (A) Radiolabeled p16 was incubated with GST-KSHV-cyclin alone (column 1), with GST-KSHV-cyclin and unphosphorylated CDK6 (column 2), or with GST-KSHV-cyclin and Thr-177 phosphorylated CDK6 (column 3). GST-KSHV-cyclin and associated proteins were precipitated by binding to glutathione-agarose, washed extensively, and the amount of bound p16 was quantified by phosphorimaging analysis following SDS-PAGE. p16 binding to unphosphorylated CDK6/GST-KSHV-cyclin (column 2) was assigned a value of 100. (B) [35S]methionine labeled KSHV-cyclin was incubated with unphosphorylated CDK6 (○), phosphorylated CDK6 (●), CDK6T177A (▵), unphosphorylated CDK6/p16 complexes (□), and phosphorylated CDK6/p16 complexes (▪). GST-CDK6 was precipitated and the amount of bound KSHV-cyclin was analyzed on SDS-PAGE gels followed by phosphorimaging. Six independent sets of data were averaged and fit to the Michaelis-Menten equation.