A Role for Mitochondrial Bak in Apoptotic Response to Anticancer Drugs

Gui Qiang Wang, Brian R. Gastman, Eva Wieckowski, Leslie A. Goldstein, Andrea Gambotto, Tae Hyoung Kim, Bingliang Fang, Asaf Rabinovitz, Xiao Ming Yin, Hannah Rabinowich

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113 Scopus citations

Abstract

In the present study a clonal Jurkat cell line deficient in expression of Bak was used to analyze the role of Bak in cytochrome c release from mitochondria. The Bak-deficient T leukemic cells were resistant to apoptosis induced by UV, staurosporin, VP-16, bleomycin, or cisplatin. In contrast to wild type Jurkat cells, these Bak-deficient cells did not respond to UV or treatment with these anticancer drugs by membranous phosphatidylserine exposure, DNA breaks, activation of caspases, or release of mitochondrial cytochrome c. The block in the apoptotic cascade was in the mitochondrial mechanism for cytochrome c release because purified mitochondria from Bak-deficient cells failed to release cytochrome c or apoptosis-inducing factor in response to recombinant Bax or truncated Bid. The resistance of Bak-deficient cells to VP-16 was reversed by transduction of the Bak gene into these cells. Also, the cytochrome c releasing capability of the Bak-deficient mitochondria was restored by insertion of recombinant Bak protein into purified mitochondria. Following mitochondrial localization, low dose recombinant Bak restored the mitochondrial release of cytochrome c in response to Bax; at increased doses it induced cytochrome c release itself. The function of Bak is independent of Bid and Bax because recombinant Bak induced cytochrome c release from mitochondria purified from Bax-/-, Bid-/-, or Bid-/- Bax-/- mice. Together, our findings suggest that Bak plays a key role in the apoptotic machinery of cytochrome c release and thus in the chemoresistance of human T leukemic cells.

Original languageEnglish (US)
Pages (from-to)34307-34317
Number of pages11
JournalJournal of Biological Chemistry
Volume276
Issue number36
DOIs
StatePublished - Sep 7 2001

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wang, G. Q., Gastman, B. R., Wieckowski, E., Goldstein, L. A., Gambotto, A., Kim, T. H., Fang, B., Rabinovitz, A., Yin, X. M., & Rabinowich, H. (2001). A Role for Mitochondrial Bak in Apoptotic Response to Anticancer Drugs. Journal of Biological Chemistry, 276(36), 34307-34317. https://doi.org/10.1074/jbc.M103526200