SAK, a new polo-like kinase, is transcriptionally repressed by p53 and induces apoptosis upon RNAi silencing

Neoplasia. 2005 Apr;7(4):312-23. doi: 10.1593/neo.04325.

Abstract

Chip profiling of a p53 temperature-sensitive tumor model identified SAK (Snk/Plk-akin kinase), encoding a new member of polo-like kinases (PLKs), as a gene strongly repressed by wild-type p53. Further characterization revealed that SAK expression was downregulated by wild-type p53 in several tumor cell models. Computer search of a 1.7-kb SAK promoter sequence revealed three putative p53 binding sites, but p53 failed to bind to any of these sites, indicating that SAK repression by p53 was not through a direct p53 binding to the promoter. Transcriptional analysis with luciferase reporters driven by SAK promoter deletion fragments identified SP-1 and CREB binding sites, which together conferred a two-fold SAK repression by p53. However, the repression was not reversed by cotransfection of SP-1 or CREB, suggesting a lack of interference between p53 and SP-1 or CREB. Significantly, p53-mediated SAK repression was largely reversed in a dose-dependent manner by Trichostatin A, a potent histone deacetylase (HDAC) inhibitor, suggesting an involvement of HDAC transcription repressors in SAK repression by p53. Biologically, SAK RNA interference (RNAi) silencing induced apoptosis, whereas SAK overexpression attenuated p53-induced apoptosis. Thus, SAK repression by p53 is likely mediated through the recruitment of HDAC repressors, and SAK repression contributes to p53-induced apoptosis.

MeSH terms

  • Amino Acid Sequence
  • Apoptosis*
  • Base Sequence
  • Binding Sites
  • Blotting, Northern
  • Cell Line, Tumor
  • Chromatin Immunoprecipitation
  • Cloning, Molecular
  • Cyclic AMP Response Element-Binding Protein / metabolism
  • DNA Fragmentation
  • Down-Regulation
  • Enzyme-Linked Immunosorbent Assay
  • Etoposide / pharmacology
  • Gene Deletion
  • Gene Expression Regulation, Neoplastic*
  • Gene Silencing*
  • Genes, Reporter
  • HeLa Cells
  • Humans
  • Luciferases / metabolism
  • Models, Genetic
  • Molecular Sequence Data
  • Oligonucleotide Array Sequence Analysis
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Protein Serine-Threonine Kinases / genetics*
  • Protein Serine-Threonine Kinases / physiology*
  • RNA Interference*
  • Software
  • Sp1 Transcription Factor / metabolism
  • Temperature
  • Time Factors
  • Transcription, Genetic*
  • Transfection
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Cyclic AMP Response Element-Binding Protein
  • Sp1 Transcription Factor
  • Tumor Suppressor Protein p53
  • Etoposide
  • Luciferases
  • PLK4 protein, human
  • Protein Serine-Threonine Kinases