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. 2005 Aug 31;95(8):644–650. doi: 10.1111/j.1349-7006.2004.tb03323.x

Reactive oxygen species (ROS) control the expression of Bcl‐2 family proteins by regulating their phosphorylation and ubiquitination

Dechao Li 1, Eisaku Ueta 1,✉, Tsuyoshi Kimura 1, Tetsuya Yamamoto 1, Tokio Osaki 1
PMCID: PMC11158795  PMID: 15298726

Abstract

We examined the influence of ROS on the phosphorylation and complex formation of Bcl‐2 family proteins in Mn‐superoxide dis‐mutase (SOD) antisense‐transfected squamous cell carcinoma cells, OSC‐4 cells. The increase of intracellular ROS level induced by cis‐diamminedichloroplatinum (CDDP) and γ‐ray treatment was greater in antisense‐transfected cells than in control vector‐trans‐fected cells, and apoptosis was more extensively induced in the former. Antisense‐transfected cells expressed high levels of Bax and Bak, but low levels of Bcl‐2 and Bcl‐XL when treated with CDDP, peplomycin, 5‐fluorouracil or γ‐rays. After treatment with these agents, the phosphorylation of protein kinase A, Bcl‐2 (Thr56) and Bad (Ser155) was increased, especially in antioxidant (N‐acetylcysteine and pyrrolidine dithiocarbamate)‐pretreated control cells, but the phosphorylation levels were very low in the antisense‐transfected cells. Bcl‐2 ubiquitination was increased, but ubiquitination of Bad and Bax was decreased in the antisense‐transfected cells, although their ubiquitination was increased by the antioxidants. These results reveal that ROS induce apoptosis by regulating the phosphorylation and ubiquitination of Bcl‐2 family proteins, resulting in increased proapoptotic protein levels and decreased antiapoptotic protein expression.

Abbreviations:

ROS

reactive oxygen species

SOD

superoxide dismutase

CDDP

cis‐diamminedichloroplatinum

5‐FU

5‐fluorouracil

PLM

peplomycin

NF‐κB

nu‐clear factor‐kappa B

BH

Bcl‐2 homology

PKA

protein kinase A

PKB

protein ki‐nase B

MAP

mitogen‐activated protein

ERK

extracellular signal‐regulated kinase

AP‐1

activator protein 1

VDAC

voltage‐dependent anion channel

NAC

N‐acetylcysteine

PDTC

pyrrolidine dithiocarbamate

DCFH‐DA

2′,7′‐dichlorofluo‐rescein diacetate

MB

mitochondrial buffer

MFI

mean fluorescence intensity

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