Characterization of calcineurin-dependent response element binding protein and its involvement in copper-metallothionein gene expression in Neurospora
- Department of Biochemistry, Osmania University, Hyderabad 500007 (India)
- Discovery Research Laboratory, Dr. Reddy's Research Foundation, Miyapur, Hyderabad (India)
- Department of Animal Sciences, University of Hyderabad, Hyderabad 500046 (India)
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 {beta}-subunit of calcineurin (Ca{sup 2+}-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.
- OSTI ID:
- 20854338
- Journal Information:
- Biochemical and Biophysical Research Communications, Vol. 345, Issue 3; Other Information: DOI: 10.1016/j.bbrc.2006.05.010; PII: S0006-291X(06)01051-5; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
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