Depletion of PKD1 by an antisense oligodeoxynucleotide induces premature G1/S-phase transition

Eur J Hum Genet. 2004 Jun;12(6):433-40. doi: 10.1038/sj.ejhg.5201136.

Abstract

Autosomal dominant polycystic kidney disease (ADPKD) is characterized by the growth of epithelial cells and the influx of cyst fluid. The 14-kb mRNA of the polycystic kidney disease gene, PKD1, encodes the polycystin-1 protein, whose function remains unknown. In this study, we observed that polycystin-1 localized in epithelial cell-cell contacts of 293 cells. We found, by bromodeoxyuridine (BrdU) incorporation experiments and Western blot analysis of S-phase-specific cyclins, that the depletion of polycystin-1 led to an increased cell proliferation rate and caused a premature G1/S-phase transition. In addition, we showed that the depletion of polycystin-1 reduced the amount of p53 in 293 cells irradiated by UV light, suggesting that polycystin-1 acts as a regulator of G1 checkpoint, which controls entry into the S phase and prevents the replication of damaged DNA. Our results might provide an insight into the formation and progression of ADPKD cysts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Western
  • Bromodeoxyuridine
  • Cell Adhesion / radiation effects
  • Cell Proliferation / radiation effects*
  • Cells, Cultured
  • DNA Damage / radiation effects
  • DNA Replication
  • Epithelial Cells / metabolism
  • Epithelial Cells / pathology
  • Epithelial Cells / radiation effects
  • G1 Phase / physiology*
  • G1 Phase / radiation effects
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Oligodeoxyribonucleotides, Antisense / pharmacology*
  • Polycystic Kidney, Autosomal Dominant / genetics
  • Polycystic Kidney, Autosomal Dominant / metabolism
  • Proteins / antagonists & inhibitors*
  • Proteins / genetics*
  • Proteins / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • S Phase / physiology*
  • S Phase / radiation effects
  • TRPP Cation Channels
  • Tumor Suppressor Protein p53 / metabolism
  • Ultraviolet Rays

Substances

  • Oligodeoxyribonucleotides, Antisense
  • Proteins
  • RNA, Messenger
  • TRPP Cation Channels
  • Tumor Suppressor Protein p53
  • polycystic kidney disease 1 protein
  • Bromodeoxyuridine