The death substrate Gas2 binds m-calpain and increases susceptibility to p53-dependent apoptosis

R Benetti, G Del Sal, M Monte, G Paroni, C Brancolini, C Schneider

Research output: Contribution to journalArticle

Abstract

Gas2 is a caspase-3 substrate that plays a role in regulating microfilament and cell shape changes during apoptosis. Here we provide evidence that overexpression of Gas2 efficiently increases cell susceptibility to apoptosis following UV irradiation, etoposide and methyl methanesulfonate treatments, and that these effects are dependent on increased p53 stability and transcription activity. To investigate possible pathways linking Gas2 to p53, a yeast two-hybrid screen swas performed, indicating m-calpain as a strong Gas2- interacting protein. Moreover, we demonstrate that Gas2 physically interacts with m-calpain in vivo and that recombinant Gas2 inhibits calpain-dependent processing of p53. Importantly, the Gas2 dominant-negative form (Gas2171-314) that binds calpain but is unable to inhibit its activity abrogates Gas2's ability to stabilize p53, to enhance p53 transcriptional activity and to induce p53-dependent apoptosis. Finally, we show that Gas2 is able to regulate the levels of p53 independently of Mdm2 status, suggesting that, like calpastatin, it may enhance p53 stability by inhibiting calpain activity.

Original languageEnglish
Pages (from-to)2702-14
Number of pages13
JournalEMBO Journal
Volume20
Issue number11
DOIs
Publication statusPublished - Jun 1 2001

Fingerprint

Calpain
Apoptosis
Substrates
Methyl Methanesulfonate
Cell Shape
Etoposide
Transcription
Actin Cytoskeleton
Caspase 3
Yeast
Yeasts
Irradiation
Processing
m-calpain
Proteins

Keywords

  • Animals
  • Apoptosis/drug effects
  • Calpain/genetics
  • Cell Line
  • Cell Survival
  • Etoposide/pharmacology
  • Genes, Reporter
  • Humans
  • Luciferases/genetics
  • Methyl Methanesulfonate/pharmacology
  • Mice
  • Mice, Inbred BALB C
  • Microfilament Proteins/metabolism
  • Osteosarcoma
  • Recombinant Proteins/metabolism
  • Saccharomyces cerevisiae
  • Sequence Deletion
  • Transcription, Genetic
  • Transfection
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53/metabolism
  • Ultraviolet Rays

Cite this

The death substrate Gas2 binds m-calpain and increases susceptibility to p53-dependent apoptosis. / Benetti, R; Del Sal, G; Monte, M; Paroni, G; Brancolini, C; Schneider, C.

In: EMBO Journal, Vol. 20, No. 11, 01.06.2001, p. 2702-14.

Research output: Contribution to journalArticle

Benetti, R ; Del Sal, G ; Monte, M ; Paroni, G ; Brancolini, C ; Schneider, C. / The death substrate Gas2 binds m-calpain and increases susceptibility to p53-dependent apoptosis. In: EMBO Journal. 2001 ; Vol. 20, No. 11. pp. 2702-14.
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AU - Schneider, C

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AB - Gas2 is a caspase-3 substrate that plays a role in regulating microfilament and cell shape changes during apoptosis. Here we provide evidence that overexpression of Gas2 efficiently increases cell susceptibility to apoptosis following UV irradiation, etoposide and methyl methanesulfonate treatments, and that these effects are dependent on increased p53 stability and transcription activity. To investigate possible pathways linking Gas2 to p53, a yeast two-hybrid screen swas performed, indicating m-calpain as a strong Gas2- interacting protein. Moreover, we demonstrate that Gas2 physically interacts with m-calpain in vivo and that recombinant Gas2 inhibits calpain-dependent processing of p53. Importantly, the Gas2 dominant-negative form (Gas2171-314) that binds calpain but is unable to inhibit its activity abrogates Gas2's ability to stabilize p53, to enhance p53 transcriptional activity and to induce p53-dependent apoptosis. Finally, we show that Gas2 is able to regulate the levels of p53 independently of Mdm2 status, suggesting that, like calpastatin, it may enhance p53 stability by inhibiting calpain activity.

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