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Query: UNIPROT:P06889 (Mol)
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We have studied the effect of membrane fatty acid composition on replicative DNA synthetic activity in mitochondria isolated from Saccharomyces cerevisiae. Cells containing different levels of membrane unsaturated fatty acids were obtained by growth of a fatty acid desaturase mutant of Saccharomyces cerevisiae in glucose-limited chemostat cultures supplemented with various concentrations of Tween 80. Arrhenius plots of DNA synthetic activity in isolated mitochondria show a discrete discontinuity at specific temperature which are dependent on the membrane unsaturated fatty acid content of the mitochondria. This indicates a functional association of DNA replication with the mitochondrial membrane in Saccharomyces cerevisiae.
Mol Biol Rep 1975 Jul
PMID:Evidence for a functional association of DNA synthesis with the membrane in mitochondria of Saccharomyces cerevisiae. 109 37

The Arabidopsis FAD7 gene encodes a chloroplast omega-3 fatty acid desaturase that catalyzes the desaturation of lipid-linked dienoic fatty acids (18:2 and 16:2). An 825 bp FAD7 promoter fragment upstream from the transcriptional start point contained several short sequences which were homologous to the cis-elements (box II, G-box, etc.) conserved in many light-responsive genes. We introduced the FAD7 promoter fused to the beta-glucuronidase (GUS) or the luciferase (LUC) reporter gene into tobacco plants. The -825 promoter sequence conferred tissue-specific and light-responsive expression to both these reporter genes in transgenic tobacco, indicating that these expressions of the FAD7 gene were regulated mainly at the transcriptional level. Histochemical GUS staining showed that the activity of the FAD7 promoter is restricted to the tissues with chloroplast-containing cells although the staining was noticeably absent in the chloroplast-containing cells associated with vascular systems. The 5' deletion experiments of the promoter revealed that the -362/-166 region, containing two putative box II sequences, was responsible for the tissue-specific and light-responsive expression of the FAD7 gene.
Plant Mol Biol 1995 Nov
PMID:Tissue-specific and light-responsive regulation of the promoter region of the Arabidopsis thaliana chloroplast omega-3 fatty acid desaturase gene (FAD7). 853 55

Lipids of axenically-cultured Giardia lamblia trophozoites were compared with those of cells undergoing in vitro encystation. Although the lipid composition of the organisms grossly resembled those of low-bile or high-bile culture media, differences were clearly detected. Encysting trophozoites incubated in a high-bile medium for 24 h had a higher concentration of unsaturated fatty acids in the total cellular lipids than did nonencysting trophozoites. The organism, but not the medium, contained linoleate and linolenate, suggesting that G. lamblia desaturates oleate. The presence of a fatty acid desaturase activity in the organism was demonstrated by the conversion of a radiolabeled monounsaturated fatty acid (oleate) to radiolabeled polyunsaturated fatty acids. Triglycerides, a common form of storage lipids, were unusually low in G. lamblia, but steryl esters (which can also serve as reserves) were abundant. Steryl esters increased during encystation of G. lamblia. The changes observed in G. lamblia lipids (increased fatty acid unsaturation and the accumulation of storage lipids) are consistent with parasite differentiation into a cyst stage that is able to survive outside the host at reduced temperatures and reduced available nutrient resources. This study also demonstrated that G. lamblia not only has the capacity to de novo synthesize isoprenoid lipids (ubiquinone, prenylated proteins), but it can also metabolize fatty acids by the addition of double bonds.
Mol Biochem Parasitol 1996 Oct 18
PMID:Changes in lipid composition during in vitro encystation and fatty acid desaturase activity of Giardia lamblia. 889 2

The omega-3 fatty acid desaturases are membrane-bound enzymes catalyzing the conversion of linoleic acid to linolenic acid in lipids, and are located both in the microsome and plastid envelopes as two different isoforms. A cDNA encoding the microsome omega-3 fatty acid desaturase (OsFAD3) and the corresponding genomic clone were isolated from rice (Oryza sativa L.). The OsFAD3 gene was composed of 8 exons and 7 introns. A microsatellite was present in the second exon of the OsFAD3 gene, showing polymorphism between Indica and Japonica rice varieties. The mapping of this microsatellite showed that the OsFAD3 gene was located on chromosome 11. Expression of the OsFAD3 cDNA in tobacco hairy root tissues and subsequent analysis of fatty acid compositions demonstrated the activity of the microsome omega-3 fatty acid desaturase. The OsFAD3 mRNA was abundant in root tissues, but was hardly detectable in leaves. In root tissues, a high level of the OsFAD3 mRNA was observed at 15 degrees C and 20 degrees C, with its level decreasing markedly at temperatures below 10 degrees C. The accumulation of the OsFAD3 mRNA in leaf tissues remained at quite low levels, both at normal growth temperatures and at chilling temperatures. Similar temperature responses of the OsFAD3 gene were observed both in chilling- tolerant and in chilling-intolerant rice cultivars.
Plant Mol Biol 1997 Feb
PMID:Structure, chromosomal location and expression of a rice gene encoding the microsome omega-3 fatty acid desaturase. 904 69

A cluster of genes involved in the production of phaseolotoxin, a phytotoxin produced by Pseudomonas syringae pv. phaseolicola, contains eight (phtA through phtH) complementation groups (Y. X. Zhang, K. B. Rowley, and S. S. Patil, J. Bacteriol., 175:6451-6458, 1993). In this study, sequencing of the region encompassing the phtE locus revealed six putative open reading frames (ORFs), each preceded by a putative ribosomal binding site, and all oriented in the same direction. Reverse transcription-polymerase chain reaction suggested that the phtE locus is transcribed as one large (6.4 kb) transcript, indicating that the ORFs constitute an operon. Primer extension analysis showed that the transcript begins at a T, located 31 bp upstream of the ATG codon of ORF1. Comparison of the sequences of the putative ORFs with the sequences of known genes revealed that ORF3, encoding a protein containing 395 amino acids, has 55% similarity to the acetylornithine aminotransferase gene from Escherichia coli, and the ornithine aminotransferase genes from other organisms. A lysine residue that is a binding site for pyridoxal phosphate and an arginine residue that is a binding site for the alpha-carboxylate group of the substrate are conserved in ORF3. These data suggest that ORF3 encodes a protein involved in the biosynthesis of ornithine, a constituent of phaseolotoxin. ORF5, encoding a peptide of 378 amino acid residues, possesses a helix-turn-helix motif at the C-terminal end that is characteristic of the AraC family of transcriptional factors, and there is a possible leucine zipper at the N-terminal end of this peptide. ORF6, encoding a protein of 327 amino acids, has about 40% similarity with the fatty acid desaturase gene, desA, of Synechocystis Pcc6803 and considerable similarity with fatty acid desaturase genes from other organisms. ORF6 and desA show very similar hydropathy profiles and both contain a copper binding signature. Computer searches did not discover significant homologies in the data base for the other ORFs, but hydropathy analysis showed that all of them contain one to several hydrophobic domains, suggesting that the gene products of these ORFs may be membrane associated.
Mol Plant Microbe Interact 1997 Nov
PMID:The phtE locus in the phaseolotoxin gene cluster has ORFs with homologies to genes encoding amino acid transferases, the AraC family of transcriptional factors, and fatty acid desaturases. 935 42

We have isolated two maize cDNAs and the corresponding genes encoding fatty acid desaturase with Arabidopsis thaliana FAD7 gene as a probe. They shared almost 90% identity at DNA sequence level. Northern analysis revealed that both genes are expressed in leaves, but not in roots at normal temperature- and low temperature-growth condition. The overall level of these transcripts are elevated upon exposure to low temperature. The tissue-specific expression and DNA sequence data indicate that both genes encode plastidic omega-3 fatty acid desaturases. One of them is expressed exclusively at normal temperature but not at 5 degrees C, whereas the other is expressed inversely. We, therefore, termed them ZmFAD7 and ZmFAD8, respectively. Among other stresses, high-salt treatment induced the accumulation of the ZmFAD7 and ZmFAD8 transcripts in roots but drought had no effect on their expression. Cycloheximide induced the accumulation of the ZmFAD7 transcript in roots. The genomic clones of ZmFAD7 and ZmFAD8 consist of 8 exons and 7 introns as same as in the cases of A. thaliana FAD7 and FAD8 genes and the sizes of the 6 internal exons were identical among them. A phylogenetic analysis of ZmFAD7, ZmFAD8 amino acid sequences and those originated from other plant species is also presented.
Plant Mol Biol 1998 Jan
PMID:Two maize genes encoding omega-3 fatty acid desaturase and their differential expression to temperature. 948 41

Unsaturated fatty acids are essential lipid components of Mucor rouxii. Gamma-linolenic acid (GLA) is synthesized via the desaturase enzymes: delta9-desaturase catalyzes mono-unsaturated fatty acids that are utilized as substrate for GLA biosynthesis. We cloned and characterized a M. rouxii gene highly homologous to delta9-desaturase genes. This sequence encodes for a protein of 452 amino acids and contains two introns of 60 and 61 nucleotides. Delta9-desaturase of M. rouxii is expressed during cell growth when cells are subjected to temperature shifts. At 30 degrees C, the mRNA level of late log phase is about 6.4-fold higher than that of early log phase. A shift from 30 to 15 degrees C induced transcription of delta9-desaturase gene in both early and late log phases. However, the pattern of increased transcription by cold induction varied depending on growth conditions: transcription of late log phase is higher than that of early log phase. These results indicate that cell growth and low temperature influence the expression of delta9-desaturase gene and fatty acid composition of M. rouxii.
Mol Cell Biol Res Commun 1999 Apr
PMID:Mucor rouxii delta9-desaturase gene is transcriptionally regulated during cell growth and by low temperature. 1032 75

We report the characterisation of two cytochrome b5 genes and their spatial and temporal patterns of expression during development in olive, Olea europaea. A PCR-generated probe, based on a tobacco cytochrome b5 sequence, was used to isolate two full-length cDNA clones (cytochrome b5-15 and cytochrome b5-38) from a library derived from 13 WAF olive fruits. The cDNAs encoded proteins of 17.0 and 17.7 kDa, which contained all the characteristic motifs of cytochromes b5 from other organisms and exhibited 63% identity and 85% similarity with each other. The olive cytochrome b5-15 cDNA was then used as a probe for more detailed analysis. Southern blotting revealed a gene family of at least 4-6 members while northern blotting and in situ hybridisation showed a highly specific pattern of gene expression. Very low levels of cytochrome b5 mRNA were detected in tissues characterised by high rates of lipid accumulation, such as young expanding leaves, maturing seeds and ripening mesocarp. The cytochrome b5 genes were not induced at 6 degrees C and their response to ABA was relatively slow compared with fatty acid desaturase genes. In contrast, high levels of cytochrome b5 gene expression were found in young fruits at the pattern formation (globular/heart) stage of embryogenesis and in vascular and transmitting tissues of male and female reproductive organs. The data are consistent with a major role for cytochrome b5 in developmental processes related to plant reproduction in addition to being an electron donor to microsomal desaturases.
Plant Mol Biol 1999 May
PMID:Temporal and spatial gene expression of cytochrome B5 during flower and fruit development in olives. 1039 47

A clone for a plastid omega-3 fatty acid desaturase (FAD) was isolated from a leaf cDNA library of hot pepper (Capsicum annuum L.). The nucleotide sequence of a 1,317 bp open reading frame in the CachFAD showed 80.9% homology with that of tobacco plant. It codes for a polypeptide of 438 amino acids with molecular mass of 50.5 kDa and a pI of 8.14. The CachFAD had a putative transit peptide for targeting the chloroplast. Genomic Southern hybridization indicated that it exists as small gene family. Northern hybridization revealed that its mRNA was present in leaves, but not in roots. Transcript levels in the leaves upon wounding increased rapidly to reach the first peak between 1-3 h, decreased thereafter and slightly increased at 24 h after wounding. The levels of linolenic acid (18:3) in wounded leaves also reached the first peak at 6 h, decreased thereafter and reached the second peak at 18 h. These results indicated that wounding not only enhanced the accumulation of the CachFAD mRNA but also increased the conversion of linoleic acid (18:2) to linolenic acid (18:3) in leaf lipids of hot pepper.
Mol Cells 2000 Oct 31
PMID:CDNA cloning of chloroplast omega-3 fatty acid desaturase from Capsicum annuum and its expression upon wounding. 1110 Nov 38

In our attempt to understand the cold shock response of Bacillus subtilis, we report on the role of the B. subtilis fatty acid desaturase (FA-D) Des during membrane adaptation to low temperatures and demonstrate its importance during cold shock. A des null mutant was constructed and analysed in comparison with its parental strain. Growth studies and large-scale comparative fatty acid (FA) analysis revealed a severe cold-sensitive phenotype of the des deletion mutant during the absence of isoleucine and showed that four unsaturated fatty acid (UFA) species differing in length, branching pattern and position of the double bond are synthesized in B. subtilis JH642 but not in the des null mutant. Apart from the lack of UFA synthesis, the FA-D deletion strain showed a dramatically altered saturated fatty acid (SFA) profile at the onset of the stationary growth phase in the presence of exogenous isoleucine sources. Expression of des integrated in trans at the amyE locus of the des deletion strain not only cured the cold-sensitive phenotype observed for the des mutant but allowed much better growth than in strain JH642 after a shift from 37 degrees C to 15 degrees C. These results show that, during cold shock adaptation, des expression can completely replace the isoleucine-dependent, long-term, FA branching adaptation mechanism. We conclude that the crucial aspect in cold adaptation of the cytoplasmic membrane is not its specific molecular composition but rather its physical status in terms of its fluidity.
Mol Microbiol 2001 Mar
PMID:Role of the Bacillus subtilis fatty acid desaturase in membrane adaptation during cold shock. 1125 47


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