PTEN M-CBR3, a versatile and selective regulator of inositol 1,3,4,5,6-pentakisphosphate (Ins(1,3,4,5,6)P5). Evidence for Ins(1,3,4,5,6)P5 as a proliferative signal

Elaine A Orchiston, Deborah Bennett, Nick R Leslie, Rosemary G Clarke, Lucinda Winward, C Peter Downes, Stephen T Safrany

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The PTEN (phosphatase and tensin homologue deleted on chromosome 10) tumor suppressor is a phosphatidylinositol 3,4,5-trisphosphate (PtdInsP3) 3-phosphatase that plays a crucial role in regulating many cellular processes by antagonizing the phosphoinositide 3-kinase signaling pathway. Although able to metabolize soluble inositol phosphates in vitro, the question of their significance as physiological substrates is unresolved. We show that inositol phosphates are not regulated by wild type PTEN, but that a synthetic mutant, PTEN M-CBR3, previously thought to be inactive toward inositides, can selectively regulate inositol 1,3,4,5,6-pentakisphosphate (Ins(1,3,4,5,6)P5). Transfection of U87-MG cells with PTEN M-CBR3 lowered Ins(1,3,4,5,6)P5 levels by 60% without detectable effect on PtdInsP3. Although PTEN M-CBR3 is a 3-phosphatase, levels of myo-inositol 1,4,5,6-tetrakisphosphate were not increased, whereas myo-inositol 1,3,4,6-tetrakisphospate levels increased by 80%. We have used PTEN M-CBR3 to study the physiological function of Ins(1,3,4,5,6)P5 and have found that Ins(1,3,4,5,6)P5 does not modulate PKB phosphorylation, nor does it regulate clathrin-mediated epidermal growth factor receptor internalization. By contrast, PTEN M-CBR3 expression, and the subsequent lowering of Ins(1,3,4,5,6)P5, are associated with reduced anchorage-independent colony formation and anchorage-dependent proliferation in U87-MG cells. Our results, together with previously published data, suggest that Ins(1,3,4,5,6)P5 has a role in proliferation.
Original languageEnglish
Pages (from-to)1116-1122
Number of pages7
JournalJournal of Biological Chemistry
Volume279
Issue number2
DOIs
Publication statusPublished - 9 Jan 2004

Keywords

  • Animals
  • Blotting, Western
  • Cell Cycle
  • Cell Division
  • Cell Line
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Polyacrylamide Gel
  • Epidermal Growth Factor
  • Flow Cytometry
  • Gene Expression Regulation, Enzymologic
  • HeLa Cells
  • Humans
  • Inositol Phosphates
  • Lipid Metabolism
  • Mice
  • Mutation
  • NIH 3T3 Cells
  • PTEN Phosphohydrolase
  • Phosphatidylinositol 3-Kinases
  • Phosphoric Monoester Hydrolases
  • Phosphorylation
  • Precipitin Tests
  • Receptor, Epidermal Growth Factor
  • Signal Transduction
  • Time Factors
  • Transfection
  • Tumor Suppressor Proteins

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