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Andreia Carina Turchetto-Zolet, Ana Paula Christoff, Franceli Rodrigues Kulcheski, Guilherme Loss-Morais, Rogerio Margis, Marcia Margis-Pinheiro
Since the first diacylglycerol acyltransferase (DGAT) gene was characterized in plants, a number of studies have focused on understanding the role of DGAT activity in plant triacylglycerol (TAG) biosynthesis. DGAT enzyme is essential in controlling TAGs synthesis and is encoded by different genes. DGAT1 and DGAT2 are the two major types of DGATs and have been well characterized in many plants. On the other hand, the DGAT3 and WS/DGAT have received less attention. In this study, we present the first general view of the presence of putative DGAT3 and WS/DGAT in several plant species and report on the diversity and evolution of these genes and its relationships with the two main DGAT genes (DGAT1 and DGAT2)...
October 3, 2016: Genetics and Molecular Biology
Zhi-Hui Zhang, Xiao-Qian Liu, Cheng Zhang, Wei He, Hua Wang, Yuan-Hua Chen, Xiao-Jing Liu, Xi Chen, De-Xiang Xu
Previous studies demonstrated that acute alcohol intoxication caused hepatic lipid accumulation. The present study showed that acute alcohol intoxication caused hepatic lipid accumulation in Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic sterol-regulatory element binding protein (SREBP)-1, a transcription factor regulating fatty acid and triglyceride (TG) synthesis, was activated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic Fas, Acc, Scd-1 and Dgat-2, the key genes for fatty acid and TG synthesis, were up-regulated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice...
2016: Scientific Reports
Tomoko Hatanaka, William Serson, Runzhi Li, Paul Armstrong, Keshun Yu, Todd Pfeiffer, Xi-Le Li, David Hildebrand
Increasing the production of plant oils such as soybean oil as a renewable resource for food and fuel is valuable. Successful breeding for higher oil levels in soybean, however, usually results in reduced protein, a second valuable seed component. This study shows that by manipulating a highly active acyl-CoA:diacylglycerol acyltransferase (DGAT) the hydrocarbon flux to oil in oilseeds can be increased without reducing the protein component. Compared to other plant DGATs, a DGAT from Vernonia galamensis (VgDGAT1A) produces much higher oil synthesis and accumulation activity in yeast, insect cells, and soybean...
September 28, 2016: Journal of Agricultural and Food Chemistry
Ángela Rojas, Jose A Del Campo, Sophie Clement, Matthieu Lemasson, Marta García-Valdecasas, Antonio Gil-Gómez, Isidora Ranchal, Birke Bartosch, Juan D Bautista, Arielle R Rosenberg, Francesco Negro, Manuel Romero-Gómez
Quercetin is a natural flavonoid, which has been shown to have anti hepatitis C virus (HCV) properties. However, the exact mechanisms whereby quercetin impacts the HCV life cycle are not fully understood. We assessed the effect of quercetin on different steps of the HCV life cycle in Huh-7.5 cells and primary human hepatocytes (PHH) infected with HCVcc. In both cell types, quercetin significantly decreased i) the viral genome replication; ii) the production of infectious HCV particles and iii) the specific infectivity of the newly produced viral particles (by 85% and 92%, Huh7...
2016: Scientific Reports
Curtis Brandt, Pamela J McFie, Scot J Stone
Acyl-CoA:1,2-diacylglycerol acyltransferase (DGAT)-2 is one of the two DGAT enzymes that catalyzes the synthesis of triacylglycerol, which is an important form of stored energy for eukaryotic organisms. There is currently limited information available regarding how DGAT2 and triacylglycerol synthesis are regulated. Recent studies have indicated that DGAT2 can be regulated by changes in gene expression. How DGAT2 is regulated post-transcriptionally remains less clear. In this study, we demonstrated that DGAT2 is a very unstable protein and is rapidly degraded in an ubiquitin-dependent manner via the proteasome...
October 15, 2016: Biochemical Journal
Peter Gajdoš, Rodrigo Ledesma-Amaro, Jean-Marc Nicaud, Milan Čertík, Tristan Rossignol
In the oleaginous yeast Yarrowia lipolytica, the diacylglycerol acyltransferases (DGATs) are major factors for triacylglycerol (TAG) synthesis. The Q4 strain, in which the four acyltransferases have been deleted, is unable to accumulate lipids and to form lipid bodies (LBs). However, the expression of a single acyltransferase in this strain restores TAG accumulation and LB formation. Using this system, it becomes possible to characterize the activity and specificity of an individual DGAT. Here, we examined the effects of DGAT overexpression on lipid accumulation and LB formation in Y...
September 2016: FEMS Yeast Research
Annika Röttig, Carl Simon Strittmatter, Jennifer Schauer, Sebastian Hiessl, Anja Poehlein, Rolf Daniel, Alexander Steinbüchel
UNLABELLED: Recently, we isolated a novel Streptomyces strain which can accumulate extraordinarily large amounts of triacylglycerol (TAG) and consists of 64% fatty acids (dry weight) when cultivated with glucose and 50% fatty acids (dry weight) when cultivated with cellobiose. To identify putative gene products responsible for lipid storage and cellobiose utilization, we analyzed its draft genome sequence. A single gene encoding a wax ester synthase/acyl coenzyme A (CoA):diacylglycerol acyltransferase (WS/DGAT) was identified and heterologously expressed in Escherichia coli The purified enzyme AtfG25 showed acyltransferase activity with C12- or C16-acyl-CoA, C12 to C18 alcohols, or dipalmitoyl glycerol...
October 1, 2016: Applied and Environmental Microbiology
BeiBei Chen, Junejie Wang, Gaoyang Zhang, Jiaqi Liu, Sehrish Manan, Honghong Hu, Jian Zhao
Diacylglycerol acyltransferases (DGATs) play a key role in plant triacylglycerol (TAG) biosynthesis. Two type 1 and 2 DGATs from soybean were characterized for their functions in TAG biosynthesis and physiological roles. GmDGAT1A is highly expressed in seeds while GmDGAT2D is mainly expressed in flower tissues. They showed different expression patterns in response to biotic and abiotic stresses. GmDGAT2D was up-regulated by cold and heat stress and ABA signaling, and repressed by insect biting and jasmonate, whereas GmDGAT1A show fewer responses...
June 27, 2016: Scientific Reports
Dan Meyers, Adele Noe, Atish Salunke, Aishwarya Movva, Kenneth Kulmatycki, Srikanth Neelakantham, Anne Crissey, Tapan Majumdar, Jin Chen
Pradigastat, a novel diacylglycerol acyltransferase 1 inhibitor, has been studied in familial chylomicronemia syndrome. To evaluate the effects of supra-therapeutic concentrations of pradigastat on the QTc interval, two studies were conducted. The first study assessed the safety, tolerability and pharmacokinetics of single escalating intravenous doses of pradigastat (10, 30, 100 and 115mg over 60 minutes) in healthy adults. Single IV doses were safe, well tolerated, and at the higher doses resulted in supra-therapeutic pradigastat exposure...
June 8, 2016: Clinical Pharmacology in Drug Development
Shiwei Li, Shibin Xu, Yanli Ma, Shuang Wu, Yu Feng, Qingpo Cui, Lifeng Chen, Shuang Zhou, Yuanyuan Kong, Xiaoyu Zhang, Jialei Yu, Mengdi Wu, Shaobing O Zhang
To identify genes that regulate the dynamics of lipid droplet (LD) size, we have used the genetically tractable model organism Caenorhabditis elegans, whose wild-type LD population displays a steady state of size with an upper limit of 3 μm in diameter. From a saturated forward genetic screen of 6.7 × 10(5) mutagenized haploid genomes, we isolated 118 mutants with supersized intestinal LDs often reaching 10 μm. These mutants define nine novel complementation groups, in addition to four known genes (maoc-1, dhs-28, daf-22, and prx-10)...
2016: G3: Genes—Genomes—Genetics
Alexandra K Leamy, Clinton M Hasenour, Robert A Egnatchik, Irina A Trenary, Cong-Hui Yao, Gary J Patti, Masakazu Shiota, Jamey D Young
Experiments in a variety of cell types, including hepatocytes, consistently demonstrate the acutely lipotoxic effects of saturated fatty acids, such as palmitate (PA), but not unsaturated fatty acids, such as oleate (OA). PA+OA co-treatment fully prevents PA lipotoxicity through mechanisms that are not well defined but which have been previously attributed to more efficient esterification and sequestration of PA into triglycerides (TGs) when OA is abundant. However, this hypothesis has never been directly tested by experimentally modulating the relative partitioning of PA/OA between TGs and other lipid fates in hepatocytes...
September 2016: Biochimica et Biophysica Acta
Nemanja Vujic, Jess Porter Abate, Stefanie Schlager, Tovo David, Dagmar Kratky, Suneil K Koliwad
The final step of triacylglycerol synthesis is catalyzed by acyl-CoA:diacylglycerol acyltransferases (DGATs). We have previously shown that ApoE-/-Dgat1-/- mice are protected from developing atherosclerosis in association with reduced foam cell formation. However, the role of DGAT1, specifically in myeloid and other hematopoietic cell types, in determining this protective phenotype is unknown. To address this question, we reconstituted the bone marrow of irradiated Ldlr-/-mice with that from wild-type (WT→ Ldlr-/-) and Dgat1-/-(Dgat1-/-→ Ldlr-/-) donor mice...
2016: PloS One
Keith Roesler, Bo Shen, Ericka Bermudez, Changjiang Li, Joanne Hunt, Howard G Damude, Kevin G Ripp, John D Everard, John R Booth, Leandro Castaneda, Lizhi Feng, Knut Meyer
Kinetically improved diacylglycerol acyltransferase (DGAT) variants were created to favorably alter carbon partitioning in soybean (Glycine max) seeds. Initially, variants of a type 1 DGAT from a high-oil, high-oleic acid plant seed, Corylus americana, were screened for high oil content in Saccharomyces cerevisiae Nearly all DGAT variants examined from high-oil strains had increased affinity for oleoyl-CoA, with S0.5 values decreased as much as 4.7-fold compared with the wild-type value of 0.94 µm Improved soybean DGAT variants were then designed to include amino acid substitutions observed in promising C...
June 2016: Plant Physiology
Curtis Brandt, Pamela J McFie, Scot J Stone
BACKGROUND: MGAT3 catalyzes the synthesis of 1,2-diacylglycerol from 2-monoacylglycerol in an acyl CoA-dependent reaction. Although initially identified as an MGAT enzyme, MGAT3 is more closely related to DGAT2 than to MGAT1 and MGAT2. Furthermore, MGAT3 possesses both DGAT and MGAT activities, in vitro. MGAT3 is almost exclusively expressed in the small intestine in humans, suggesting that it has a role in dietary fat absorption. Although identified many years ago, little information is available regarding the contribution of MGAT3 to triacylglycerol biosynthesis...
July 1, 2016: Biochemical and Biophysical Research Communications
Da-Wei Li, Shi-Ying Cen, Yu-Hong Liu, Srinivasan Balamurugan, Xin-Yan Zheng, Adili Alimujiang, Wei-Dong Yang, Jie-Sheng Liu, Hong-Ye Li
Oleaginous microalgae have received a considerable attention as potential biofuel feedstock. However, lack of industry-suitable strain with lipid rich biomass limits its commercial applications. Targeted engineering of lipogenic pathways represents a promising strategy to enhance the efficacy of microalgal oil production. In this study, a type 2 diacylglycerol acyltransferase (DGAT), a rate-limiting enzyme in triacylglycerol (TAG) biosynthesis, was identified and overexpressed in heterokont oleaginous microalga Nannochloropsis oceanica for the first time...
July 10, 2016: Journal of Biotechnology
Xi Wang, Xiaochao Xiong, Na Sa, Sanja Roje, Shulin Chen
With the growing attention to global warming and energy sustainability, biosynthesis of lipids by photosynthetic microorganisms has attracted more interest for the production of renewable transportation fuels. Recently, the cyanobacterium Synechocystis sp. PCC 6803 has been widely used for biofuel production through metabolic engineering because of its efficient photosynthesis and well-developed genetic tools. In lipid biosynthesis, glycerol-3-phosphate (G3P) is a key node for both CO2 fixation and lipid metabolism in cyanobacteria...
July 2016: Applied Microbiology and Biotechnology
Maria Morgan-Bathke, Liang Chen, Elisabeth Oberschneider, Debra Harteneck, Michael D Jensen
Adipose tissue fatty acid storage varies according to sex, adipose tissue depot and degree of fat gain. However, the mechanism(s) for these variations is not completely understood. We recently published findings based on the glycerol 3-phosphate acyltransferase (GPAT) enzyme activity assay we optimized for use with human adipose tissue. These findings include a decrease in total GPAT and GPAT1 as a function of adipocyte size in both omental and subcutaneous adipose tissue and a strong, positive correlations between ACS, GPAT, and DGAT activities for both sexes and depots and between these storage factors and palmitate storage rates into TAG...
January 2016: Adipocyte
Michael S Greer, Xue Pan, Randall J Weselake
Diacylglycerol acyltransferase (DGAT) catalyzes the acyl-CoA-dependent acylation of sn-1, 2-diacylglycerol to produce triacylglycerol, which is the main component of the seed oil of Brassica oilseed species. Phylogenetic analysis of the amino acid sequences encoded by four transcriptionally active DGAT1 genes from Brassica napus suggests that the gene forms diverged over time into two clades (I and II), with representative members in each genome (A and C). The majority of the amino acid sequence differences in these forms of DGAT1, however, reside outside of motifs suggested to be involved in catalysis...
June 2016: Lipids
Huiying Zhao, Dongsheng Fan, Dale R Nyholt, Yuedong Yang
Genome-wide association studies (GWAS) have indicated potential to identify heritability of common complex phenotypes, but traditional approaches have limited ability to detect hiding signals because single SNP has weak effect size accounting for only a small fraction of overall phenotypic variations. To improve the power of GWAS, methods have been developed to identify truly associated genes by jointly testing effects of all SNPs. However, equally considering all SNPs within a gene might dilute strong signals of SNPs in real functional categories...
August 2016: Human Mutation
Hua Ning, Zongxiang Sun, Yunyun Liu, Lei Liu, Liuyi Hao, Yaxin Ye, Rennan Feng, Jie Li, Ying Li, Xia Chu, Songtao Li, Changhao Sun
The detrimental role of hepatic lipotoxicity has been well-implicated in the pathogenesis of NAFLD. Previously, we reported that inhibiting autophagy aggravated saturated fatty acid (SFA)-induced hepatotoxicity. Insulin, a physiological inhibitor of autophagy, is commonly increased within NAFLD mainly caused by insulin resistance. We therefore hypothesized that insulin augments the sensitivity of hepatocyte to SFA-induced lipotoxicity. The present study was conducted via employing human and mouse hepatocytes, which were exposed to SFAs, insulin, or their combination...
2016: Nutrients
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