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Cloning and characterization of 5'-flanking region of oxalate decarboxylase gene from Flammulina velutipes

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Title Cloning and characterization of 5'-flanking region of oxalate decarboxylase gene from Flammulina velutipes
 
Creator Azam, Mohammad
Kesarwani, Meenu
Chakraborty, Subhra
Natarajan, Krishnamurthy
Datta, Asis
 
Subject low pH responsive element
oxalic acid
pH regulated promoter
 
Description The oxalate-degrading enzyme, oxalate decarboxylase (OXDC), was purified and characterized from Flammulina elutipes, a basidiomycetous fungus [Mehta and Datta (1991) J. Biol. Chem. 266, 23548–23553]. The cDNA cloning and analyses revealed that OXDC transcription was induced by oxalic acid. However, in this report, we show that OXDC transcription is induced by low pH, not by oxalate. To understand the regulatory mechanism of OXDC expression, we have cloned and analysed a 580-bp genomic fragment from the 5h-flanking region of the OXDC gene. Sequence analysis showed the presence of several eukaryotic transcription factor binding motifs within the k580 bp of the upstream region. Electrophoretic-mobility-shift assays with partially purified cell extracts revealed specific binding of a factor in acid-induced, but not in uninduced, extracts. Furthermore,
DNase I protection assays using the partially purified fraction from oxalic acid-induced extract revealed a footprint of a 13-bp sequence 5hGCGGGGTCGCCGA3h, termed low pH responsive element (LPRE), corresponding to the k287 to k275 bp region
of the OXDC promoter. Our results suggest that in F. elutipes cells, activation of OXDC transcription in response to low pH is
mediated by the binding of a novel transcription factor through
the LPRE site in the OXDC promoter.
This research project was supported by the Department of Biotechnology, Ministry
of Science and Technology, Government of India
 
Date 2013-10-17T10:20:48Z
2013-10-17T10:20:48Z
2002
 
Type Article
 
Identifier Biochemical J., 367: 67-75
http://hdl.handle.net/123456789/22
 
Language en
 
Publisher Biochemical Society