title: Analysis of the promoter region of the gene LIP1 encoding triglyceride lipase from Fusarium graminearum.
abstract: Triglyceride lipases catalyze the reversible degradation of glycerol esters with long-chain fatty acids into fatty acids and glycerol. In silico analysis of 5'-end flanking sequence of the gene LIP1 encoding a triglyceride lipase from the wheat head blight pathogen Fusarium graminearum revealed the presence of several cis-regulatory elements. To delineate the function of these regulatory elements, we constructed a series of deletion mutants in the LIP1 promoter region fused to the open reading frame of a green fluorescent protein (GFP) and assayed the promoter activity. Analysis of GFP expression levels in mutants indicated that a 563-bp promoter sequence was sufficient to drive the expression of LIP1 and regulatory elements responsible for the gene induction were located within the 563-372bp region. To further investigate the regulatory elements, putative cis-acting elements spanned within the 563-372bp region were mutated using a targeted mutagenesis approach. A CCAAT box, a CreA binding site, and a fatty acid responsive element (FARE) were identified and confirmed to be required for the basal expression of LIP1, glucose suppression and fatty acid induction, respectively.
authors.affiliation: Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, SK, Canada. jie.feng@gov.ab.ca
authors.forename: Jie
authors.initials: J
authors.lastname: Feng
authors.forename: Vijai
authors.initials: V
authors.lastname: Bhadauria
authors.forename: Guosheng
authors.initials: G
authors.lastname: Liu
authors.forename: Gopalan
authors.initials: G
authors.lastname: Selvaraj
authors.forename: Geoffrey R
authors.initials: GR
authors.lastname: Hughes
authors.forename: Yangdou
authors.initials: Y
authors.lastname: Wei
mesh_terms: D019677:Artificial Gene Fusion; D001665:Binding Sites; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008049:Lipase / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D014908:Triticum
chemical_list: D049452:Green Fluorescent Proteins; D008049:Lipase