RRC ID 36128
Author Kamthan A, Kamthan M, Kumar A, Sharma P, Ansari S, Thakur SS, Chaudhuri A, Datta A.
Title A calmodulin like EF hand protein positively regulates oxalate decarboxylase expression by interacting with E-box elements of the promoter.
Journal Sci Rep
Abstract 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 (45)Ca(2+), heat stability and Ca(2+) dependent electrophoretic shift. Hence, FvCaMLP can be considered a new addition to the category of unconventional Ca(2+) binding transcriptional regulators.
Volume 5
Pages 14578
Published 2015-10-12
DOI 10.1038/srep14578
PII srep14578
PMID 26455820
PMC PMC4600981
MeSH Amino Acid Sequence Base Sequence Binding Sites Calcium / metabolism Calmodulin / chemistry* Calmodulin / genetics Calmodulin / metabolism Carboxy-Lyases / chemistry* Carboxy-Lyases / genetics Carboxy-Lyases / metabolism EF Hand Motifs / genetics* Escherichia coli / genetics Escherichia coli / metabolism Flammulina / genetics* Flammulina / metabolism Fungal Proteins / chemistry* Fungal Proteins / genetics Fungal Proteins / metabolism Gene Expression Regulation, Fungal* Hyphae / genetics Hyphae / metabolism Models, Molecular Oxalic Acid / metabolism Protein Binding Protein Structure, Tertiary RNA, Messenger / genetics RNA, Messenger / metabolism Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / metabolism Schizosaccharomyces / genetics Schizosaccharomyces / metabolism Sequence Alignment
IF 3.998
Times Cited 8
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Yeast FYP2930(pTN54)