Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
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Reverse Transcriptional Polymerase Chain Reaction (RT-PCR), Rapid Amplification of the cDNA ends (RACE) and Thermal asymmetric interlaced (TAIL)-PCR were used successfully to clone the open reading frame (1,377 bp) of delta9-fatty acid desaturase gene (named as RAD9) and its promoter region from oil-producing fungi Rhizopus arrhizus. Functional identification of the protein was done by sub-cloning RAD9 into the expression vector pYES2.0 to generate a recombinant plasmid pYRAD9, which was then subsequently transformed into Saccharomyces cerevisiae delta9-fatty acid desaturase mutation strain L8-14C to be expressed under the control of GAL1 promoter. The transformant containing RAD9 named as L8-14C-pYRAD9 could grow on the synthetic minimal medium plate with out oleic acid supplement. Fatty acid analysis also showed that the transformant contained 16:1 and 18:1. This indicated that pYRAD9 could successfully complement the mutation of L8-14C. Computational analysis of the nucleotide sequence of RAD9 promoter showed several basic transcriptional elements including a CAAT box, a GC box, a CACCC box, two TATA boxes and also several putative target-binding sites for transcription factors, which have been reported to be involved in the regulation of lipid metabolism. Preliminary functional analysis of this promoter in S. cerevisiae was done using lacZ report gene.
Mol Biol Rep 2009 Jan
PMID:Identification of a novel delta9-fatty acid desaturase gene and its promoter from oil-producing fungus Rhizopus arrhizus. 1793 71

Altered D-glucose metabolism prevails in the soleus muscle of rats depleted in long-chain polyunsaturated omega3 fatty acids (omega3). In these animals, the prior intravenous injection of an omega3-rich medium-chain triglyceride:fish oil emulsion (omega3-FO rats), as compared to that of an omega3-poor medium-chain triglyceride:olive oil emulsion (omega3-OO rats), may either correct or aggravate selected metabolic variables. This study deals with the fatty acid pattern of soleus phospholipids and triglycerides in control animals versus omega3-depleted rats not injected with any lipid emulsion (omega3-NI rats) and in omega3-OO versus omega3-FO rats. In each group of omega3-depleted rats, age-related changes were also monitored. The omega3-depleted rats displayed low long-chain polyunsaturated omega3 fatty acid content, facilitated metabolism of long-chain polyunsaturated omega6 fatty acids, and increased Delta9-desaturase activity. Both the age-related changes in lipid variables and those attributable to the prior intravenous injection of the omega3-rich lipid emulsion consisted either in a move towards normalization or in the opposite direction, i.e. towards aggravation of the defect found in the omega3-depleted rats. Emphasis is placed, therefore, on the unusual situation found in the soleus muscle of omega3-depleted rats, in which both lipid and metabolic variables may be either favourably or adversely affected by the same environmental factor(s).
Int J Mol Med 2007 Dec
PMID:Perturbation and age-related changes in the fatty acid pattern of soleus muscle phospholipids and triglycerides in rats depleted in long-chain polyunsaturated omega3 fatty acids. 1798

Omega(3)-fatty acid desaturase and Delta(12)-fatty acid desaturase of Pichia pastoris with distinguishable regioselectivity and high degree of sequence similarity were chosen for regioselectivity research. Chimeras were constructed in which Histidine-rich boxes 1, 2 and the carboxyl terminal region of omega(3)-fatty acid desaturase were replaced with corresponding region of Delta(12)-fatty acid desaturase. The replacement was found to result in a change of regioselectivity from omegay to x + 3 by functionally characterizing these chimeric enzymes in Saccharomyces cerevisae strain INVScI. Using site-directed mutagenesis, we further demonstrated that seven conserved amino acids of omega(3)-fatty acid desaturase within the first two Histidine-rich regions are responsible for the regioselectivity switch. Therefore, the regioselectivity of fatty acid desaturases may be better understood by investigating the evolutionary relationships of different fatty acid desaturases.
Mol Biol Rep 2009 Mar
PMID:Evolution-related amino acids play important role in determining regioselectivity of fatty acid desaturase from Pichia pastoris. 1828 82

We report the first isolation and characterization of insect fatty acid Delta12-desaturase genes, AdD12Des from house cricket (Acheta domesticus) and TcD12Des from the red flour beetle (Tribolium castaneum), responsible for the production of linoleic acid from oleic acid. Sequence analysis shows the cricket and flour beetle Delta12-desaturase genes have evolved independently from all previously known Delta12-desaturases and are much more closely related to the archetypal stearoyl-Coenzyme A-acting desaturase from rat than to the phospholipid-acting Delta12-desaturases widely reported in plants. Phylogenetic and functional analysis indicates the cricket AdD12Des gene may have evolved from an ancestral Delta9-desaturase. By contrast, the beetle Delta12-desaturase is distantly related to the cricket genes and beetle Delta9-desaturases suggesting evolution by an independent route. Linoleic acid has key physiological roles in insects and this is the first report of genes capable of producing this essential fatty acid in higher animals.
Insect Mol Biol 2008 Dec
PMID:Isolation and functional characterization of two independently-evolved fatty acid Delta12-desaturase genes from insects. 1913 76

Fatty acid composition of fungi is analysed through the gas chromatography technique. With specific activity a novel enzyme Delta6-fatty acid desaturase was screened and isolated from Rhizopus nigricans. In this study R. nigricans was identified as a fungal species that produced plentiful gamma-linolenic acid. A 1,475 bp full-length cDNA, designated as RnD6D here, with high homology to fungal Delta6-fatty acid desaturase genes was isolated from R. nigricans using reverse transcription polymerase chain reaction and rapid amplification of cDNA ends methods. Sequence analysis indicated that this cDNA sequence had an open reading frame of 1,380 bp encoding a deduced polypeptide of 459 amino acids. Bioinformatics analysis characterized the putative RnD6D protein as a typical membrane-bound desaturase, including three conserved histidine-rich motifs, hydropathy profile and a cytochrome b5-like domain in the N-terminus. The corresponding genomic sequence of RnD6D was 1,689 bp with 4 introns, which was at least one intron more than other fungal Delta6-fatty acid desaturase genes. To elucidate the function of this novel putative desaturase, the coding sequence was expressed in Saccharomyces cerevisiae strain INVScl. A novel peak corresponding to gamma-linolenic acid methyl ester standards was detected with the same retention time, which was absent in the cell transformed with empty vector. The result demonstrated that the coding produced Delta6-fatty acid desaturase activity of RnD6D which led to the accumulation of gamma-linolenic acid. The functionally active RnD6D gene cloned here defined a novel Delta6-fatty acid desaturase from R. nigricans.
Mol Biol Rep 2009 Nov
PMID:Identification and characterization of a novel delta6-fatty acid desaturase gene from Rhizopus nigricans. 1915 21

Using a custom-made cDNA microarray, global transcriptional analyses were conducted to identify genes differentially regulated in the pheromone gland as compared to the remaining insect tissue of the moth Agrotis segetum (Noctuidae). A two-fold or larger difference in relative expression levels was found for 227 of 864 genes investigated comparing the two tissues. Unexpectedly, an antennal binding protein homologue, containing a pheromone-binding/general odorant-binding protein PFAM domain, was expressed at a 56-fold higher level in the pheromone gland. Relatively higher expression levels in the pheromone gland were also found for other gene representatives putatively encoding odorant-binding proteins and chemosensory proteins, as well as a number of gene representatives putatively encoding proteins involved in juvenile hormone interactions. The largest relative up-regulation (84-fold) in the pheromone gland was found for a gene encoding a Delta11-desaturase homologue implicated in desaturation of pheromone precursors. For three gene representatives, the expression patterns were independently verified by quantitative real-time PCR (qPCR). Additionally the expression pattern in the pheromone gland for the Delta11-desaturase homologue was shown by qPCR to follow the previously known pattern of pheromone production in female A. segetum, both with respect to age and circadian rhythm, whereas the expression of a Delta9-desaturase and a chemosensory protein homologue did not share this pattern.
Insect Biochem Mol Biol 2009 Jul
PMID:Global transcriptional analysis of pheromone biosynthesis-related genes in the female turnip moth, Agrotis segetum (Noctuidae) using a custom-made cDNA microarray. 1937 28

The development of the dauer form of Caenorhabditis elegans daf-2(e1370) enhances the expression of genes such as fatty acid desaturase fat-6 and fat-7 and fatty acid elongase elo-2, and increases the level of triglyceride (TAG). RNA interference (RNAi) of the fat-6, fat-7, and elo-2 genes lowers fat accumulation in the nematode. We recently clarified the fact that RNAi of fat-related genes, especially fat-2, reduced fat accumulation and activated DAF-16. FAT-2 regulates the first step of polyunsaturated fatty acid (PUFA) synthesis. RNAi of fat-2 induced nuclear translocation of DAF-16 and increased the level of TAG that could be detected by Oil Red-O, but suppressed the accumulation of lipid dyed by Nile red. TAG levels are also increased in the adult daf-2(e1370), whereas Nile red staining showed fat reduction. Introduction of RNAi of fat-2, fat-6, fat-7, and elo-2 genes into the daf-16 deficient worm recovered Nile red-stained lipid storage. These results suggest that Nile red stained the lipids except TAG, and that the levels of these lipids are regulated by daf-16. In fat-2, fat-6, fat-7, and elo-2-RNAi worms, the Nile red-stained fat level was restored through addition of fatty acids, especially PUFA. This suggests that reduction of Nile red-dyed lipid reflects the disorder of fatty acid metabolism. Furthermore, treatment of the fat-2-RNAi worm with PUFA--using the fatty acids from linoleic acid through eicosapentaenoic acid--suppressed nuclear localization of DAF-16. These results suggest that PUFA acts as a mediator of daf-2/insulin signaling and that daf-16 might be involved in fatty acid homeostasis under the control of PUFA.
Mol Cell Endocrinol 2010 Jul 29
PMID:Polyunsaturated fatty acids are involved in regulatory mechanism of fatty acid homeostasis via daf-2/insulin signaling in Caenorhabditis elegans. 2022 39

In Lake Superior there are three principal forms of lake trout (Salvelinus namaycush): lean, siscowet and humper. Wild lean and siscowet differ in the shape and relative size of the head, size of the fins, location and size of the eyes, caudal peduncle shape and lipid content of the musculature. To investigate the basis for these phenotypic differences, lean and siscowet lake trout, derived from gametes of wild populations in Lake Superior, were reared communally under identical environmental conditions for 2.5 years. Fish were analysed for growth, morphometry and lipid content, and differences in liver transcriptomics were investigated using Roche 454 GS-FLX pyrosequencing. The results demonstrate that key phenotypic differences between wild lean and siscowet lake trout such as condition factor, morphometry and lipid levels, persist in these two forms when reared in the laboratory under identical environmental conditions. This strongly suggests that these differences are genetic and not a result of environmental plasticity. Transcriptomic analysis involving the comparison of hepatic gene frequencies (RNA-seq) and expression (quantitative reverse transcription-polymerase chain reaction (qPCR)) between the two lake trout forms, indicated two primary gene groups that were differentially expressed; those involving lipid synthesis, metabolism and transport (acyl-CoA desaturase, acyl-CoA binding protein, peroxisome proliferator-activated receptor gamma, and apolipoproteins), and those involved with immunity (complement component C3, proteasome, FK506 binding protein 5 and C1q proteins). The results demonstrate that RNA-seq can be used to identify differentially expressed genes; however, some discrepancies between RNA-seq analysis and qPCR indicate that methods for deep sequencing may need to be refined and/or different RNA-seq platforms utilized.
Mol Ecol 2010 Mar
PMID:A genetic basis for the phenotypic differentiation between siscowet and lean lake trout (Salvelinus namaycush). 2033 79

The antifungal mode of action of chitosan has been studied for the last 30 years, but is still little understood. We have found that the plasma membrane forms a barrier to chitosan in chitosan-resistant but not chitosan-sensitive fungi. The plasma membranes of chitosan-sensitive fungi were shown to have more polyunsaturated fatty acids than chitosan-resistant fungi, suggesting that their permeabilization by chitosan may be dependent on membrane fluidity. A fatty acid desaturase mutant of Neurospora crassa with reduced plasma membrane fluidity exhibited increased resistance to chitosan. Steady-state fluorescence anisotropy measurements on artificial membranes showed that chitosan binds to negatively charged phospholipids that alter plasma membrane fluidity and induces membrane permeabilization, which was greatest in membranes containing more polyunsaturated lipids. Phylogenetic analysis of fungi with known sensitivity to chitosan suggests that chitosan resistance may have evolved in nematophagous and entomopathogenic fungi, which naturally encounter chitosan during infection of arthropods and nematodes. Our findings provide a method to predict the sensitivity of a fungus to chitosan based on its plasma membrane composition, and suggests a new strategy for antifungal therapy, which involves treatments that increase plasma membrane fluidity to make fungi more sensitive to fungicides such as chitosan.
Mol Microbiol 2010 Feb
PMID:Membrane fluidity determines sensitivity of filamentous fungi to chitosan. 2048 94

SUMMARY The preformed (Z,Z)-1-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15-diene (AFD) is the most active antifungal compound in avocado; it affects the quiescence of Colletotrichum gloeosporioides in unripe fruit. One of the genes encoding Delta(12) fatty acid desaturase (avfad12) was hypothesized to take part in the biosynthesis of AFD, and its expression pattern and enzymatic activity were determined in relation to the content of AFD. Using avfad12-3 as a probe, high levels of expression were detected in young fruits and leaves, where the level of AFD was highest. In contrast, Northern analysis of RNA from mature leaves and fruits showed no transcripts from the avfad12 gene family and lower AFD content. The transcripts from the avfad12 gene family, the enzymatic activity of Delta(12) fatty acid desaturase, and the level of AFD in unripe-resistant fruits increased transiently when the fruits were inoculated with C. gloeosporioides or exposed to ethylene (40 microL/L), low temperature (4 degrees C) or 1 mm H(2)O(2), but ripe fruits were not affected. The effect of H(2)O(2) on the transcripts from the avfad12 gene family is of specific importance, because reactive oxygen species were produced by unripe-resistant host fruit soon after inoculation of C. gloeosporioides. In addition, the fungus itself produced H(2)O(2) in culture medium at pH 5.0, which is similar to the pH of unripe-resistant fruit, but not at pH 7.0. Treatments that enhanced Delta(12) fatty acid desaturase activity increased the concentration of the AFD precursor, linoleic acid, and its incorporation into AFD; these treatments also caused a delay in decay development. The present results demonstrate temporal relationships among the transcripts from the avfad12 gene family, the synthesis of the precursor of AFD (linoleic acid), the AFD content and quiescence of C. gloeosporioides in unripe fruits.
Mol Plant Pathol 2004 Nov 01
PMID:Expression of Delta(12) fatty acid desaturase during the induced accumulation of the antifungal diene in avocado fruits. 2056 31


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