کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1939696 | 1050765 | 2006 | 4 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Characterization of calcineurin-dependent response element binding protein and its involvement in copper-metallothionein gene expression in Neurospora Characterization of calcineurin-dependent response element binding protein and its involvement in copper-metallothionein gene expression in Neurospora](/preview/png/1939696.png)
In continuation of our recent observations indicating the presence of a lone calcineurin-dependent response element (CDRE) in the −3730 bp upstream region of copper-induced metallothionein (CuMT) gene of Neurospora [K.S. Kumar, S. Dayananda, C. Subramanyam, Copper alone, but not oxidative stress, induces copper-metallothionein gene in Neurospora crassa, FEMS Microbiol. Lett. 242 (2005) 45–50], we isolated and characterized the CDRE-binding protein. The cloned upstream region of CuMT gene was used as the template to specifically amplify CDRE element, which was immobilized on CNBr-activated Sepharose 4B for use as the affinity matrix to purify the CDRE binding protein from nuclear extracts obtained from Neurospora cultures grown in presence of copper. Two-dimensional gel electrophoresis of the affinity purified protein revealed the presence of a single 17 kDa protein, which was identified and characterized by MALDI-TOF. Peptide mass finger printing of tryptic digests and analysis of the 17 kDa protein matched with the regulatory β-subunit of calcineurin (Ca2+-calmodulin dependent protein phosphatase). Parallel identification of nuclear localization signals in this protein by in silico analysis suggests a putative role for calcineurin in the regulation of CuMT gene expression.
Journal: Biochemical and Biophysical Research Communications - Volume 345, Issue 3, 7 July 2006, Pages 1010–1013