Proteolytic cleavage of phospholipase C-γ1 during apoptosis in Molt-4 cells

Sun Sik Bae, David K. Perry, Yong Seok Oh, Jang Hyun Choi, Sehamuddin H. Galadari, Tariq Ghayur, Sung Ho Ryu, Yusuf A. Hannun, Pann Ghill Suh

Research output: Contribution to journalArticle

Abstract

Apoptosis is a cell suicide mechanism that requires the activation of cellular death proteases for its induction. We examined whether the progress of apoptosis involves cleavage of phospholipase C-γ1 (PLC-γ1), which plays a pivotal role in mitogenic signaling pathway. Pretreatment of T leukemic Molt-4 cells with PLC inhibitors such as U-73122 or ET-18-OCH3 potentiated etoposide-induced apoptosis in these cells. PLC-γ1 was fragmented when Molt- 4 cells were treated with several apoptotic stimuli such as etoposide, ceramides, and tumor necrosis factor α. Cleavage of PLC-γ1 was blocked by overexpression of Bcl-2 and by specific inhibitors of caspases such as Z- DEVD-CH2F and YVAD-cmk. Purified caspase-3 and caspase-7, group II caspases, cleaved PLC-γ1 in vitro and generated a cleavage product of the same size as that observed in vivo, suggesting that PLC-γ1 is cleaved by group II caspases in vivo. From point mutagenesis studies, Ala-Glu-Pro-Asp770 was identified to be a cleavage site within PLC-γ1. Epidermal growth factor receptor (EGFR) -induced tyrosine phosphorylation of PLC-γ1 resulted in resistance to cleavage by caspase-3 in vitro. Furthermore, cleaved PLC-γ1 could not be tyrosine-phosphorylated by EGFR in vitro. In addition, tyrosine- phosphorylated PLC-γ1 was not significantly cleaved during etoposide-induced apoptosis in Molt-4 cells. This suggests that the growth factor-induced tyrosine phosphorylation may suppress apoptosis-induced fragmentation of PLC- γ1. We provide evidence for the biochemical relationship between PLC-γ1- mediated signal pathway and apoptotic signal pathway, indicating that the defect of PLC-γ1-mediated signaling pathway can facilitate an apoptotic progression.

Original languageEnglish (US)
Pages (from-to)1083-1092
Number of pages10
JournalFASEB Journal
Volume14
Issue number9
DOIs
StatePublished - 2000

Keywords

  • PLC-γ1
  • Proteolysis
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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    Bae, S. S., Perry, D. K., Oh, Y. S., Choi, J. H., Galadari, S. H., Ghayur, T., Ryu, S. H., Hannun, Y. A., & Suh, P. G. (2000). Proteolytic cleavage of phospholipase C-γ1 during apoptosis in Molt-4 cells. FASEB Journal, 14(9), 1083-1092. https://doi.org/10.1096/fasebj.14.9.1083