Silencing of CDK2, but not CDK1, separates mitogenic from anti-apoptotic signaling, sensitizing p53 defective cells for synthetic lethality.
Small molecule inhibitors targeting CDK1/CDK2 have been clinically proven effective against a variety of tumors, albeit at the cost of profound off target toxicities. To separate potential therapeutic from toxic effects, we selectively knocked down CDK1 or CDK2 in p53 mutated HACAT cells by siRNA silencing. Using dynamic, cell cycle wide proteome arrays, we observed minor changes in overall abundance of proteins critically involved in cell cycle transition despite profound G2/M or G1/S arrest, respectively. Employing phospho site specific analyses, we identified uncoupled mitogenic, yet pro-apoptotic signaling from counter balancing anti-apoptotic activity in CDK2 disrupted cells. Moreover, a crucial role of CDK2 activity in early serum response was observed, extending well-established roles of CDKs outside their cell cycle regulating functions. In contrast, disruption of CDK1 only marginally affected phosphorylation events of crucial signaling nodes prior to G2/S transition. The data presented here suggest that the temporal separation of pro- and anti-apoptotic pathways by selective inhibition of CDK2 disrupts coherent signaling modules and may synergize with anti-proliferative drugs, averting toxic side effects from CDK1 inhibition.
a Department of Internal Medicine II , Julius-Maximilians University , Wuerzburg , Germany.
Tatyana S
TS
Nekova
b Physiological Chemistry I, Biocenter, Julius-Maximilians University , Wuerzburg , Germany.
Susanne
S
Kneitz
a Department of Internal Medicine II , Julius-Maximilians University , Wuerzburg , Germany.
Hermann
H
Einsele
c Cancer Comprehensive Center Mainfranken, Julius-Maximilians University , Wuerzburg , Germany.
Ralf
R
Bargou
a Department of Internal Medicine II , Julius-Maximilians University , Wuerzburg , Germany.|d DKD Helios Klinik Wiesbaden , Wiesbaden , Germany.
Gernot
G
Stuhler
D017209:Apoptosis; D016203:CDC2 Protein Kinase / Q000172:deficiency / Q000378:metabolism*; D002460:Cell Line; D020868:Gene Silencing; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D007700:Kinetics; D008938:Mitosis / Q000235:genetics; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction; D000072020:Synthetic Lethal Mutations / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D016203:CDC2 Protein Kinase
CDK1; CDK2; HACAT; cell cycle; p53; phosphorylation; serine-threonine kinase; signaling pathway; tyrosine kinase