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J. Biol. Chem., Vol. 276, Issue 2, 1335-1344, January 12, 2001
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From the The neurotransmitter dopamine (DA) induces
apoptosis via its oxidative metabolites. This study shows that
glutaredoxin 2 (Grx2) from Escherichia coli and human
glutaredoxin could protect cerebellar granule neurons from DA-induced
apoptosis. E. coli Grx2, which catalyzes
glutathione-disulfide oxidoreduction via its -Cys-Pro-Tyr-Cys- active
site, penetrates into cerebellar granule neurons and exerts its
activity via NF-
Glutaredoxin Protects Cerebellar Granule Neurons from
Dopamine-induced Apoptosis by Activating NF-
B via Ref-1*
,
,
Department of Neurobiochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Tel
Aviv 69978 Israel, the § Department of Biochemistry and
Biophysics, Medical Nobel Institute for Biochemistry, Karolinska
Institute, S-171 77 Stockholm, Sweden, the ¶ Department of
Neurology and Felsenstein Medical Research Institute, Rabin Medical
Center and Sackler School of Medicine, Tel Aviv University, Ramat Aviv,
Tel Aviv 69978 Israel, and the Interdepartmental Core Facility,
Sackler School of Medicine, Tel Aviv University, Ramat Aviv,
Tel Aviv 69978 Israel
B activation. Analysis of single and double cysteine
to serine substitutions in the active site of Grx2 showed that both
cysteine residues were essential for activity. Although DA
significantly reduced NF-B binding activity, Grx2 could stimulate the binding of NF-B to DNA by: (i) translocating NF-B from the cytoplasm to the nucleus after promoting the phosphorylation and degradation of I-B
, and (ii) activating the binding of pre
existing nuclear NF-B. The DNA binding activity of NF-B itself
was essential for neuronal survival. Overexpression of I-B dominant
negative gene (I-B-
N) in granule neurons significantly reduced
their viability, irrespective of the presence of Grx2. Ref-1 expression was down-regulated by DA but up-regulated by Grx2, while treatment of
neurons with Ref-1 antisense oligonucleotide reduced the ability of
Grx2 to activate NF-B binding activity. These results show that Grx2
exerts its anti apoptotic activity through the activation of Ref-1,
which then activates NF-B.
*
This work was supported in part by grants from the Israeli
Academy of Sciences and Humanities and the Israeli Ministry of Health,
by Swedish Cancer Society Grant 961, and by Swedish Medical Research
Council Grant 03XS-13005-01A (to A. H. and A. V.-G.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Neurobiochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel. Tel.: 972-3-6409782; Fax: 972-3-6407643; E-mail: barzilai@post.tau.ac.il.
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