The expression level of the voltage-dependent anion channel controls life and death of the cell.
Mitochondria not only generate cellular energy, but also act as the point for cellular decisions leading to apoptosis. The voltage-dependent anion channel (VDAC), as a major mitochondrial outer-membrane transporter, has an important role in energy production by controlling metabolite traffic and is also recognized as a key protein in mitochondria-mediated apoptosis. In this study, the role of VDAC1 in regulating cell survival and death was investigated by silencing endogenous human (h)VDAC1 expression by using a short hairpin RNA (shRNA)-expressing vector. The shRNA effectively down-regulated the expression in human T-REx-293 cells of hVDAC1 but not murine (m)VDAC1. Cells in which hVDAC1 expression was decreased by approximately 90% proliferated extremely slowly. Normal growth was, however, restored upon expression of mVDAC1 in a tetracycline-regulated manner. Although low tetracycline concentrations promoted cell growth, high concentrations induced mVDAC1 overexpression, leading to cell death. Cells with low levels of VDAC1 showed 4-fold-lower ATP-synthesis capacity and contained low ATP and ADP levels, with a strong correlation between ATP levels and cell growth, suggesting limited metabolite exchange between mitochondria and cytosol. The possibility of suppressing endogenous hVDAC1 expression and introducing native and mutated mVDAC1 is used to further explore the involvement of VDAC1 in apoptosis. Cells suppressed for hVDAC1 but expressing either native mVDAC1 or an E72Q mutant underwent apoptosis induced by various stimuli that can be inhibited by ruthenium red in the native cells but not in the mutated cells, suggesting that VDAC1 regulates apoptosis independent of the apoptosis-inducing pathway.
Department of Life Sciences, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Salah
S
Abu-Hamad
Sara
S
Sivan
Varda
V
Shoshan-Barmatz
D000255:Adenosine Triphosphate / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology; D001483:Base Sequence; D016923:Cell Death / Q000502:physiology*; D002468:Cell Physiological Phenomena; D002470:Cell Survival / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D034622:RNA Interference; D016337:RNA, Catalytic / Q000235:genetics; D050995:Voltage-Dependent Anion Channel 1 / Q000235:genetics*
D016337:RNA, Catalytic; C418481:hairpin ribozyme; D000255:Adenosine Triphosphate; D050995:Voltage-Dependent Anion Channel 1