Molecular cloning and characterization of the human p19(INK4d) gene promoter

FEBS Lett. 2002 Apr 24;517(1-3):272-6. doi: 10.1016/s0014-5793(02)02647-9.

Abstract

p19(INK4d), a member of the INK4 family of cyclin-dependent kinase (CDK) inhibitors, negatively regulates the cyclin D-CDK4/6 complexes, which promote G1/S transition by phosphorylating the retinoblastoma tumor-suppressor gene product. To investigate the mechanism of transcriptional regulation of the p19(INK4d) gene, we characterized the 5'-flanking region of the human p19(INK4d) gene. The cap-site hunting method revealed that the transcription starts at -16 nucleotide (nt) upstream of the initiation codon. The 5'-flanking region of the human p19(INK4d) gene was ligated to a luciferase reporter gene and possessed functional promoter activity. Luciferase assay with a series of truncated 5'-flanking regions indicated that the region from -81 to -2 nt could drive the transcription of the p19(INK4d) gene. Several Sp1 and activating protein 2 binding sites are located within the region from -81 to -2 nt. Mutation of the second Sp1 binding site from -33 to -25 nt decreased the promoter activity. Collectively, it was demonstrated that the human p19(INK4d) gene is under the control of TATA-less promoter and the Sp1 binding site is involved in the transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Flanking Region / genetics*
  • Base Sequence
  • Binding Sites
  • Cell Cycle Proteins*
  • Cloning, Molecular
  • Cyclin-Dependent Kinase Inhibitor p16 / genetics*
  • Cyclin-Dependent Kinase Inhibitor p19
  • DNA-Binding Proteins / genetics
  • Genes, Reporter
  • Humans
  • Molecular Sequence Data
  • Mutagenesis
  • Promoter Regions, Genetic*
  • Sequence Analysis
  • Sequence Deletion
  • Sp1 Transcription Factor / genetics*
  • Transcription Factor AP-2
  • Transcription Factors / genetics
  • Transcription Initiation Site
  • Transcription, Genetic*
  • Tumor Cells, Cultured

Substances

  • CDKN2D protein, human
  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p16
  • Cyclin-Dependent Kinase Inhibitor p19
  • DNA-Binding Proteins
  • Sp1 Transcription Factor
  • Transcription Factor AP-2
  • Transcription Factors