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A calmodulin like EF hand protein positively regulates oxalate decarboxylase expression by interacting with E-box elements of the promoter

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Title A calmodulin like EF hand protein positively regulates oxalate decarboxylase expression by interacting with E-box elements of the promoter
 
Creator Kamthan, Ayushi
Kamthan, Mohan
Kumar, Avinash
Sharma, Pratima
Ansari, Sekhu
Thakur, Sarjeet Singh
Chaudhuri, Abira
Datta, Asis
 
Subject Fungal biology
Transcriptional
regulatory elements
Fungal biology
RNAi
 
Description Accepted date: 03 September 2015
Oxalate decarboxylase (OXDC) enzyme has immense biotechnological applications due to its ability to decompose anti-nutrient oxalic acid. Flammulina velutipes, an edible wood rotting fungus responds to oxalic acid by induction of OXDC to maintain steady levels of pH and oxalate anions outside the fungal hyphae. Here, we report that upon oxalic acid induction, a calmodulin (CaM) like protein-FvCaMLP, interacts with the OXDC promoter to regulate its expression. Electrophoretic mobility shift assay showed that FvCamlp specifically binds to two non-canonical E-box elements (AACGTG) in the OXDC promoter. Moreover, substitutions of amino acids in the EF hand motifs resulted in loss of DNA binding ability of FvCamlp. F. velutipes mycelia treated with synthetic siRNAs designed against FvCaMLP showed significant reduction in FvCaMLP as well as OXDC transcript pointing towards positive nature of the regulation. FvCaMLP is different from other known EF hand proteins. It shows sequence similarity to both CaMs and myosin regulatory light chain (Cdc4), but has properties typical of a calmodulin, like binding of 45Ca2+, heat stability and Ca2+ dependent electrophoretic shift. Hence, FvCaMLP can be considered a new addition to the category of unconventional Ca2+ binding transcriptional regulators.
We thank Dr. Taro Nakamura (Osaka city university, Japan) for the plasmid pTN54; confocal facility,
NIPGR for immunoflourescence microscopy. AK, MK and AKu thank CSIR for fellowship. AD thanks
Department of Biotechnology, Government of India, for funding the project.
 
Date 2016-01-06T09:26:51Z
2016-01-06T09:26:51Z
2015
 
Type Article
 
Identifier Scientific Reports, 5: 14578
2045-2322
http://172.16.0.77:8080/jspui/handle/123456789/523
http://www.nature.com/articles/srep14578
10.1038/srep14578
 
Language en_US
 
Publisher Nature Publishing Group