Extended Data Fig. 7: Differences between CDK12 and other CDKs highlight the residues involved in CR8-induced recruitment of DDB1.
From: The CDK inhibitor CR8 acts as a molecular glue degrader that depletes cyclin K
a, Sequence alignment of CDK12 and CDK13. b, Sequence alignment of CDK12 and CDK9. c, Multiple sequence alignment of different human CDKs. In a–c, asterisks denote contacts with CR8 and circles indicate contacts with DDB1 (coloured according to DDB1 domains; see Fig. 2). Arrows mark differences at the DDB1–CR8–CDK interface. d, Titration of CDK12–cyclin KAlexa488 (0–3.75 μM) into 50 nM DDB1terbium and 10 μM CR8 or DMSO (n = 3). ‘No DDB1’ only contains terbium-coupled streptavidin and shows concentration-dependent fluorophore effects. e, Titration of CDK13–cyclin KAlexa488 (0–3.75 μM) into 50 nM DDB1terbium and 10 μM CR8 or DMSO (n = 3). f, Titration of CDK9–cyclin KAlexa488 (0–3.75 μM) into 50 nM DDB1terbium and 10 μM CR8 or DMSO (n = 3). g, CUL4NEDD8–RBX1–DDB1 in vitro ubiquitination of cyclin K bound to CDK12, CDK13 or CDK9 (n = 2). h, Titration of CDK12(L1033A/W1036A)–cyclin KAlexa488 (0–3.75 μM) into 50 nM DDB1terbium and 10 μM CR8 or DMSO (n = 3). i, Titration of CDK12(ΔCTE)–cyclin KAlexa488 (0–3.75 μM) into 50 nM DDB1terbium and 10 μM CR8 or DMSO (n = 3). CDK12(ΔCTE) is a truncated version of CDK12 (amino acids 713–1032).
