GABPA

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GABPA

A gene on chromosome 21q21.3 that encodes the alpha subunit of the GA-binding protein, which forms a tetrameric complex with two beta subunits and stimulates transcription of target genes. The encoded protein may downregulate cytochrome oxidase expression and may play a role in nuclear control of mitochondrial function.
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For the overexpression of zebrafish nrf2a, capped nrf2a RNA was synthesized from pCS2nrf2 [11] using an SP6 mMESSAGE mMACHINE in vitro transcription kit (Ambion, Austin, TX).
In vitro synthesized mRNA of nrf2a, the functional ortholog of mammalian Nrf2 in zebrafish, was injected into 1-cell stage of zebrafish embryos, and the gene expression in the injected embryos at 8h after the injection was examined using 16KMZH chips, which contain 16,399 probes [19].
Overexpression of nrf2a in mRNA-injected embryos was confirmed by a real-time qPCR (Figure S1, 75.5-fold higher compared to uninjected embryos).
Furthermore, looking back on the lineup of genes that were upregulated by the overexpression of nrf2a (Table S3), two more related genes were found: taldo1 (transaldolase 1) and fbp1a (fructose-1,6-bisphosphatase 1a).
The result indicated that the expression levels of pgd andfbp1a (1.40- and 2.76-fold, resp.), but not pckl, pcxb, and taldol (1.21-, 0.75-, and 0.70-fold, resp.), were significantly upregulated by the overexpression of nrf2a (Figure 3(b)).
Weak DEM-induced expression of fbp1a was also observed in the gills, but this induction was independent of nrf2a genotypes.
The genes that were selected included gclm (glutamate-cysteine ligase, modifier subunit), gsto2 (glutathione S-transferase omega 2), gsr (glutathione reductase), sqstml (sequestosome 1), and keapla (kelch-like ECH-associated protein 1a), together with a well-studied nrf2a target, gstp1 (glutathione S-transferase pi 1).
In our microarray analysis, we also found 55 genes that were downregulated by the overexpression of nrf2a (Table S4).
In the present microarray analysis, the cut-off value for significant upregulation by the overexpression of nrf2a was set at a 1.5-fold change instead of a 2-fold change in the previous studies [19, 20].
(a) The gene expression of the indicated proteasome subunits in 8 h postfertilization (hpf) wild-type embryos injected with or without 100 pg of nrf2a mRNA at the 1- cell stage was analyzed by a real-time qPCR.
(b) The gene expression of the indicated enzymes related to glucose metabolism in 8 hpf wild-type embryos injected with or without 100 pg of nrf2a mRNA at the 1-cell stage was analyzed by a real-time qPCR.