CDK12-mediated transcriptional regulation of noncanonical NF-κB components is essential for signaling

Sci Signal. 2018 Jul 31;11(541):eaam8216. doi: 10.1126/scisignal.aam8216.

Abstract

Members of the family of nuclear factor κB (NF-κB) transcription factors are critical for multiple cellular processes, including regulating innate and adaptive immune responses, cell proliferation, and cell survival. Canonical NF-κB complexes are retained in the cytoplasm by the inhibitory protein IκBα, whereas noncanonical NF-κB complexes are retained by p100. Although activation of canonical NF-κB signaling through the IκBα kinase complex is well studied, few regulators of the NF-κB-inducing kinase (NIK)-dependent processing of noncanonical p100 to p52 and the subsequent nuclear translocation of p52 have been identified. We discovered a role for cyclin-dependent kinase 12 (CDK12) in transcriptionally regulating the noncanonical NF-κB pathway. High-content phenotypic screening identified the compound 919278 as a specific inhibitor of the lymphotoxin β receptor (LTβR), and tumor necrosis factor (TNF) receptor superfamily member 12A (FN14)-dependent nuclear translocation of p52, but not of the TNF-α receptor-mediated nuclear translocation of p65. Chemoproteomics identified CDK12 as the target of 919278. CDK12 inhibition by 919278, the CDK inhibitor THZ1, or siRNA-mediated knockdown resulted in similar global transcriptional changes and prevented the LTβR- and FN14-dependent expression of MAP3K14 (which encodes NIK) as well as NIK accumulation by reducing phosphorylation of the carboxyl-terminal domain of RNA polymerase II. By coupling a phenotypic screen with chemoproteomics, we identified a pathway for the activation of the noncanonical NF-κB pathway that could serve as a therapeutic target in autoimmunity and cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Bone Neoplasms / drug therapy
  • Bone Neoplasms / metabolism
  • Bone Neoplasms / pathology
  • Cyclin-Dependent Kinases / antagonists & inhibitors
  • Cyclin-Dependent Kinases / genetics
  • Cyclin-Dependent Kinases / metabolism*
  • Cyclins / genetics
  • Cyclins / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic*
  • High-Throughput Screening Assays
  • Humans
  • Indoles / pharmacology
  • Lymphotoxin beta Receptor / antagonists & inhibitors
  • Lymphotoxin beta Receptor / genetics
  • Lymphotoxin beta Receptor / metabolism
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • NF-kappa B p52 Subunit / genetics
  • NF-kappa B p52 Subunit / metabolism
  • NF-kappaB-Inducing Kinase
  • Osteosarcoma / drug therapy
  • Osteosarcoma / metabolism*
  • Osteosarcoma / pathology
  • Propionates / pharmacology
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Proteome
  • Signal Transduction
  • TWEAK Receptor / antagonists & inhibitors
  • TWEAK Receptor / genetics
  • TWEAK Receptor / metabolism
  • Tumor Cells, Cultured

Substances

  • Antineoplastic Agents
  • CCNK protein, human
  • Cyclins
  • Indoles
  • LTBR protein, human
  • Lymphotoxin beta Receptor
  • NF-kappa B
  • NF-kappa B p52 Subunit
  • NFKB2 protein, human
  • Propionates
  • Proteome
  • TNFRSF12A protein, human
  • TWEAK Receptor
  • Protein Serine-Threonine Kinases
  • CDK12 protein, human
  • Cyclin-Dependent Kinases