scholarly journals A novel redox-sensing transcriptional regulator CyeR controls expression of an Old Yellow Enzyme family protein in Corynebacterium glutamicum

Microbiology ◽  
2010 ◽  
Vol 156 (5) ◽  
pp. 1335-1341 ◽  
Author(s):  
Shigeki Ehira ◽  
Haruhiko Teramoto ◽  
Masayuki Inui ◽  
Hideaki Yukawa
Keyword(s):  
Oxidative Stress ◽  
Dna Binding ◽  
Corynebacterium Glutamicum ◽  
Redox Regulation ◽  
Transcriptional Regulator ◽  
Binding Activity ◽  
Old Yellow Enzyme ◽  
Dna Binding Activity ◽  
Family Protein ◽  
Redox Sensing

Corynebacterium glutamicum cgR_2930 (cyeR) encodes a transcriptional regulator of the ArsR family. Its gene product, CyeR, was shown here to repress the expression of cyeR and the cgR_2931 (cye1)–cgR_2932 operon, which is located upstream of cyeR in the opposite orientation. The cye1 gene encodes an Old Yellow Enzyme family protein, members of which have been implicated in the oxidative stress response. CyeR binds to the intergenic region between cyeR and cye1. Expression of cyeR and cye1 is induced by oxidative stress, and the DNA-binding activity of CyeR is impaired by oxidants such as diamide and H2O2. CyeR contains two cysteine residues, Cys-36 and Cys-43. Whereas mutation of the former (C36A) has no effect on the redox regulation of CyeR activity, mutating the latter (C43A, C43S) abolishes the DNA-binding activity of CyeR. Cys-43 of CyeR and its C36A derivative are modified upon treatment with diamide, suggesting an important role for Cys-43 in the redox regulation of CyeR activity. It is concluded that CyeR is a redox-sensing transcriptional regulator that controls cye1 expression.

scholarly journals Redox Regulation of GA-binding Protein-α DNA Binding Activity

1996 ◽  
Vol 271 (41) ◽  
pp. 25617-25623 ◽  
Author(s):  
Mark E. Martin ◽  
Yurii Chinenov ◽  
Mi Yu ◽  
Tonya K. Schmidt ◽  
Xiu-Ying Yang
Keyword(s):  
Dna Binding ◽  
Redox Regulation ◽  
Binding Protein ◽  
Binding Activity ◽  
Dna Binding Activity ◽  
Ga Binding Protein

scholarly journals Redox regulation of DNA binding activity of DREF (DNA replication-related element binding factor) in Drosophila

2004 ◽  
Vol 378 (3) ◽  
pp. 833-838 ◽  
Author(s):  
Tae-Yeong CHOI ◽  
S. Young PARK ◽  
Ho-Sung KANG ◽  
Jae-Hun CHEONG ◽  
Han-Do KIM ◽  
...  
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Redox State ◽  
Redox Regulation ◽  
Binding Activity ◽  
Dna Binding Activity ◽  
Cell Extracts ◽  
Binding Factor ◽  
Related Element ◽  
Intracellular Redox

DREF [DRE (DNA replication-related element) binding factor] is an 80 kDa polypeptide homodimer which plays an important role in regulating cell proliferation-related genes. Both DNA binding and dimer formation activities are associated with residues 16–115 of the N-terminal region. However, the mechanisms by which DREF dimerization and DNA binding are regulated remain unknown. Here, we report that the DNA binding activity of DREF is regulated by a redox mechanism, and that the cysteine residues are involved in this regulation. Electrophoretic mobility shift analysis using Drosophila Kc cell extracts or recombinant DREF proteins indicated that the DNA binding domain is sufficient for redox regulation. Site-directed mutagenesis and transient transfection assays showed that Cys59 and/or Cys62 are critical both for DNA binding and for redox regulation, whereas Cys91 is dispensable. In addition, experiments using Kc cells indicated that the DNA binding activity and function of DREF are affected by the intracellular redox state. These findings give insight into the exact nature of DREF function in the regulation of target genes by the intracellular redox state.

Calorie restriction attenuates inflammatory responses to myocardial ischemia-reperfusion injury

2001 ◽  
Vol 280 (5) ◽  
pp. H2094-H2102 ◽  
Author(s):  
B. Chandrasekar ◽  
J. F. Nelson ◽  
J. T. Colston ◽  
G. L. Freeman
Keyword(s):  
Oxidative Stress ◽  
Dna Binding ◽  
Antioxidant Enzyme ◽  
Calorie Restriction ◽  
Inflammatory Responses ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Binding Activity ◽  
Dna Binding Activity ◽  
Myocardial Oxidative Stress

The life-prolonging effects of calorie restriction (CR) may be due to reduced damage from cumulative oxidative stress. Our goal was to determine the long-term effects of moderate dietary CR on the myocardial response to reperfusion after a single episode of sublethal ischemia. Male Fisher 344 rats were fed either an ad libitum (AL) or CR (40% less calories) diet. At age 12 mo the animals were anaesthetized and subjected to thoracotomy and a 15-min left-anterior descending coronary artery occlusion. The hearts were reperfused for various periods. GSH and GSSG levels, nuclear factor-κB (NF-κB) DNA binding activity, cytokine, and antioxidant enzyme expression were assessed in the ischemic zones. Sham-operated animals served as controls. Compared with the AL diet, chronic CR limited oxidative stress as seen by rapid recovery in GSH levels in previously ischemic myocardium. CR reduced DNA binding activity of NF-κB. The κB-responsive cytokines interleukin-1β and tumor necrosis factor-α were transiently expressed in the CR group but persisted longer in the AL group. Furthermore, expression of manganese superoxide dismutase, a key antioxidant enzyme, was significantly delayed in the AL group. Collectively these data indicate that CR significantly attenuates myocardial oxidative stress and the postischemic inflammatory response.

scholarly journals Redox Regulation of Adenovirus-Induced AP-1 Activation by Overexpression of Manganese-Containing Superoxide Dismutase

2002 ◽  
Vol 76 (1) ◽  
pp. 355-363 ◽  
Author(s):  
Hannah J. Zhang ◽  
Victoria J. Drake ◽  
Linjing Xu ◽  
Jianfang Hu ◽  
Frederick E. Domann ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Dna Binding ◽  
Redox Regulation ◽  
Recombinant Adenovirus ◽  
Redox Status ◽  
Binding Activity ◽  
Dna Binding Activity ◽  
Promising Tool ◽  
Gel Shift Assay ◽  
Intracellular Redox Status

ABSTRACT Adenovirus gene therapy is a promising tool in the clinical treatment of many genetic and acquired diseases. However, it has also caused pathogenic effects in organs such as the liver. The redox-sensitive transcription factors AP-1 and NF-κB have been implicated in these effects. To study the mechanisms of adenovirus-mediated AP-1 and NF-κB activation and the possible involvement of oxidative stress in adenovirus transduction, rats were injected with either replication-defective recombinant adenovirus with DNA containing the cytomegalovirus promoter region only (AdCMV), adenovirus containing human manganese-containing superoxide dismutase (MnSOD) cDNA (AdMnSOD), or vehicle. Compared to vehicle and AdCMV transduction, MnSOD gene transfer yielded a fivefold increase in liver MnSOD activity 7 days postinjection. Gel shift assay showed that AdCMV transduction induced DNA binding activity for AP-1 but not NF-κB. MnSOD overexpression abolished this activation. Western blotting analysis of c-Fos and c-Jun suggested that up-regulation of c-fos and c-jun gene expression does not directly contribute to the induction of AP-1 activation. Glutathione/glutathione disulfide ratios were decreased by adenovirus transduction and restored by MnSOD overexpression. The AP-1 binding activity that was induced by AdCMV was decreased by immunoprecipitation of Ref-1 protein. Ref-1 involvement was confirmed by restoration of AP-1 binding activity after the immunoprecipitated Ref-1 protein had been added back. AP-1 DNA binding activity was also elevated in control and AdMnSOD-injected rats after addition of the immunoprecipitated Ref-1 protein. These data indicate that cellular transduction by recombinant adenovirus stimulates AP-1 DNA binding activity. Furthermore, our results suggest that MnSOD overexpression decreases AP-1 DNA binding activity by regulating intracellular redox status, with the possible involvement of Ref-1 in this redox-sensitive pathway.

Cooperative DNA binding of the human HoxB5 (Hox-2.1) protein is under redox regulation in vitro

1993 ◽  
Vol 13 (8) ◽  
pp. 4609-4617
Author(s):  
C K Galang ◽  
C A Hauser
Keyword(s):  
Dna Binding ◽  
Redox Regulation ◽  
Mutational Analysis ◽  
Cysteine Residue ◽  
Binding Activity ◽  
Nf Kappa B ◽  
Amino Terminal ◽  
Dna Binding Activity ◽  
Specific Oxidation

The human HoxB5 (Hox-2.1) gene product is a sequence-specific DNA binding protein. Cooperative interactions stabilize in vitro DNA binding of the HoxB5 protein to tandem binding sites by at least 100-fold relative to binding to a single site. The HoxB5 homeodomain is sufficient for sequence-specific DNA binding but not for cooperative DNA binding. Here we report that the additional protein sequence required for cooperativity is a small domain adjacent to the homeodomain on the amino-terminal side. We further show that cooperative DNA binding is under redox regulation. The HoxB5 protein binds to DNA in vitro both when oxidized or reduced but binds cooperatively only when oxidized. Mutational analysis has revealed that the cysteine residue in the turn between homeodomain helices 2 and 3 is necessary for cooperative binding and redox regulation. The enhanced DNA binding of oxidized HoxB5 protein is the opposite of the redox regulation reported for other mammalian transcription factors such as Fos, Jun, USF, NF-kappa B, c-Myb, and v-Rel, in which oxidation of cysteine residues inhibits DNA binding. Thus, specific oxidation of nuclear proteins is a potential regulatory mechanism that can act to either decrease or increase their DNA binding activity.

scholarly journals The RxxRxRxxC motif conserved in all Rel/kappa B proteins is essential for the DNA-binding activity and redox regulation of the v-Rel oncoprotein.

1992 ◽  
Vol 12 (7) ◽  
pp. 3094-3106 ◽  
Author(s):  
S Kumar ◽  
A B Rabson ◽  
C Gélinas
Keyword(s):  
Dna Binding ◽  
Transcriptional Activation ◽  
Redox Regulation ◽  
Biological Activities ◽  
Cysteine Residue ◽  
Binding Activity ◽  
Lymphoid Cells ◽  
Redox Control ◽  
Dna Binding Activity

The v- and c-Rel oncoproteins bind to oligonucleotides containing kappa B motifs, form heterodimers with other members of the Rel family, and modulate expression of genes linked to kappa B motifs. Here, we report that the RxxRxRxxC motif conserved in all Rel/kappa B family proteins is absolutely required for v-Rel protein-DNA contact and its resulting transforming activity. We also demonstrate that serine substitution of the cysteine residue conserved within this motif enables v-Rel to escape redox control, thereby promoting overall DNA binding. These mutant proteins retained the ability to competitively inhibit kappa B-mediated transcriptional activation of the human immunodeficiency virus long terminal repeat but failed to efficiently transform chicken lymphoid cells both in vitro and in vivo. Our data indicate that reduction of the conserved cysteine residue in the RxxRxRxxC motif may be required for optimal DNA-protein interactions. These results provide direct biochemical evidence that the DNA-binding activity of v-Rel is subject to redox control and that the conserved cysteine residue in the RxxRxRxxC motif is critical for this regulation. These studies suggest that the DNA-binding, transcriptional, and biological activities of Rel family proteins may also be subject to redox control in vivo.

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