• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1563-1575
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• 2023

Acyl-coenzyme A (CoA):diacylglycerol acyltransferase 2 (DGAT2) catalyzes the last stage of triacylglycerol (TAG) synthesis, a process that forms ester bonds with diacylglycerols (DAG) and fatty acyl-CoA substrates. The enzymatic role of Dgat2 has been studied in various biological species. Still, the full description of how Dgat2 channels fatty acids in skeletal myocytes and the consequence thereof in glucose uptake have yet to be well established. Therefore, this study explored the mediating role of Dgat2 in glucose uptake and fatty acid partitioning under short interfering ribonucleic acid (siRNA)-mediated Dgat2 knockdown conditions. Cells transfected with Dgat2 siRNA downregulated glucose transporter type 4 (Glut4) messenger RNA (mRNA) expression and decreased the cellular uptake of [1-¹⁴C]-labeled 2-deoxyglucose up to 24.3% (p < 0.05). Suppression of Dgat2 deteriorated insulin-induced Akt phosphorylation. Dgat2 siRNA reduced [1-¹⁴C]-labeled oleic acid incorporation into TAG, but increased the level of [1-¹⁴C]-labeled free fatty acids at 3 h after initial fatty acid loading. In an experiment of chasing radioisotope-labeled fatty acids, Dgat2 suppression augmented the level of cellular free fatty acids. It decreased the level of re-esterification of free fatty acids to TAG by 67.6% during the chase period, and the remaining pulses of phospholipids and cholesteryl esters were decreased by 34.5% and 61%, respectively. Incorporating labeled fatty acids into beta-oxidation products increased in Dgat2 siRNA transfected cells without gene expression involving fatty acid oxidation. These results indicate that Dgat2 has regulatory function in glucose uptake, possibly through the reaction of TAG with endogenously released or recycled fatty acids.

• Journal of the Korean Data and Information Science Society
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• 제26권6호
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• pp.1249-1258
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• 2015

젖소의 유생산 형질에 가장 큰 영향을 미치는 유전자들 중 하나로 알려진 DGAT1 유전자의 효과를 국내 젖소 종축의 고밀도 유전체 정보를 이용하여 검증하기 위하여 본 연구를 수행하였다. 국내 젖소 씨수소로 구성된 353두의 고밀도 유전체 정보, 혈통, 추정 육종가 및 신뢰도 정보를 수집하였으며, 단일염기다형성 효과를 추정하기 위한 종속변량으로 가장 정확한 유전체 육종가를 예측할 수 있는 DeRegressed EBV를 산출하여 분석에 이용하였다. BovineSNP50 v2 패널을 이용하여 구명한 고밀도 유전자형 정보 중 유효성검증 과정을 통하여 41,051개 SNP을 선정하였으며, 각 단일 염기다형성의 실제적 유전체 육종가 기여도를 확인하기 위하여 유전체 선발방법 중 하나인 베이즈B (pi=0.99) 방법을 이용하여 SNP 효과를 추정하였다. 1메가 베이스페어의 구간으로 구성된 유전체 전장의 2,516개 윈도우 별 유전분산 설명력을 계산한 결과 상위 1, 3 윈도우가 DGAT1유전자 주변에서 발견되었으며, 이 두 윈도우의 유전분산 설명력은 각각 0.51% 및 0.48%인 것으로 나타났다. DGAT1유전자는 유전체 선발에 상업적으로 이용되는 50k SNP chip에 포함되어있지 않기 때문에 직접적인 유전자의 효과가 명확하게 드러나지는 않지만 DGAT1 유전자에 인접한 단일염기다형성들간의 연관불평형에 의하여 주변 윈도우에서 가장 높은 유전분산 설명력을 보이는 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권7호
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• pp.943-954
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• 2009

Diacylglycerol acyltransferase (DGAT) is a key enzyme that catalyzes the final and rate-limiting step of triglyceride synthesis. Both DGAT1 and DGAT2 genes code proteins with DGAT activity. Studies have shown DGAT1 polymorphisms associate with intramuscular fat deposition in beef cattle, but fewer associations between DGAT2 and beef cattle economic traits have been reported. The objective of this study was to investigate single nucleotide polymorphism (SNP) in intron3 of bovine DGAT2 and evaluate the associations of that with carcass, meat quality, and fat yield traits. Test animals were 157 commercial feedlot steers belonging to 3 Chinese native breeds (22 for Luxi, 24 for Jinnan, and 23 for Qinchuan), 3 cross populations (20 for Charolais${\times}$Fuzhou, 18 for Limousin ${\times}$Luxi, and 17 for Simmental${\times}$Jinan) and 1 Taurus pure breed population (16 Angus steers). In the current study, 15 SNP were discovered in intron3 and exon4 of DGAT2 at positions 65, 128, 178, 210, 241, 255, 270, 312, 328, 334, 365, 366, 371, 415, and 437 (named as their positions in PCR amplified fragments). Only 7 of them (128, 178, 241, 270, 312, 328, and 371) were analyzed, because SNP in three groups (65-128-255, 178-210-365 and 241-334-366) were in complete linkage disequilibrium within the group, and SNP 415 was a deletion and 437 was a null mutation. Frequencies for rare alleles in the 3 native breed populations were higher than in the 3 cross populations for 178 (p = 0.04), 270 (p = 0.001), 312 (p = 0.03) and 371 (p = 0.002). A general linear model was used to evaluate the associations between either SNP genotypes or allele substitutions and the measured traits. Results showed that SNP 270 had a significant association with the fat yield associated with kidney, pelvic cavity, heart, intestine, and stomach (KPHISY). Animals with genotype CC and CT for 270 had less (CC: -7.71${\pm}$3.3 kg and CT: -5.34${\pm}$2.5 kg) KPHISY than animals with genotype TT (p = 0.02). Allele C for 270 was associated with an increase of -4.26${\pm}$1.52 kg KPHISY (p = 0.006) and $-0.92{\pm}0.45%$ of retail cuts weight percentage (NMP, Retail cuts weight/slaughter body weight) (p = 0.045); allele G for 312 was associated with an increase of -5.45${\pm}$2.41 kg KPHISY (p = 0.026). An initial conclusion was that associations do exist between DGAT2 gene and carcass fat traits. Because of the small sample size of this study, it is proposed that further effort is required to validate these findings in larger populations.

• 농업과학연구
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• 제36권2호
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• pp.167-177
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• 2009

본 연구는 한국형 Holstein종 젖소집단의 DGAT1 유전자의 특성을 구명하고, DGAT1의 유전적 다형과 산유형질인 유량 및 유지량 간의 연관성을 구명하여 젖소집단의 유전적 개량을 위한 분자유전학적 접목을 위하여 실험을 실시하였다. Holstein종의 genomic DNA로부터 PCR기법을 이용하여 DGAT1 유전자좌를 specific primers로 증폭한 후, 1.5% agarose gel에 전기영동한 결과 411 bp의 단편이 양호하게 증폭되었음을 확인하였다. DGAT1 유전자의 염기서열을 분석한 결과 DGAT1 Q 대립유전자의 216-218 bp의 염기서열이 AUG(lysine, K)였으나, 동위치의 대립유전자 q의 염기서열은 GCG(alanine, A)로 치환되어 있음을 확인할 수 있었다. 한편 한국 Holstein종과 NCBI에서 보고된 bovine DGAT1 유전자 단편의 염기서열간에는 100% 상동성을 보였다. 한국 Holstein종 유우집단의 DGAT1 유전자형의 분포는 DGAT1 QQ, Qq 및 qq 유전자 분포가 각각 16.43, 36.43 및 47.14%로 qq 유전자형빈도가 다른 유전자형에 비하여 높았으며, 유전자빈도는 DGAT1 Q 및 q 빈도가 각각 0.35 및 0.65로 q의 빈도가 높았다. DGAT1 유전자형과 산유량인 유량 및 유지량간의 연관성에 있어서는 DGAT1 유전자형이 유량 및 유지량에서 유의적인 차이 (P<0.05)를 보였으며, DGAT1 Qq 유전자형이 QQ 및 qq 유전자형에 비하여 유량과 유지량에서 유의적인 차이(P<0.05)로 높은 수치를 나타냈다.

• Archives of Pharmacal Research
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• 제28권2호
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• pp.164-168
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• 2005

The EtOAc extract of Youngia koidzumiana significantly inhibited the diacylglycerol acyltransferase (DGAT) from rat liver microsomes. Bioactivity-guided fractionation led to the isolation of nine compounds, the structures of which were established using physicochemical and spectral data. Of the isolated compounds, oleanolic acid (2), methyl ursolate (7) and corosolic aicd (8) inhibited DGAT, with $IC_{50}$ values of 31.7, 26.4, and $44.3{\mu}M$, respectively. However, sesquit-erpenoids showed only weak inhibitory effects toward DGAT.

• Journal of Animal Science and Technology
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• 제50권3호
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• pp.293-300
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• 2008

비육 기간 중 근육 내 지방 축적 및 관련 유전자 발현 양상을 확인하기 위하여, 등심 내 지방 및 중성지방 함량을 측정하고 중성지방 생합성 관련 유전자인 ACSS2, GPAT, MGAT 및 DGAT의 유전자 발현량을 real-time PCR법을 통하여 분석하였다. 근육 내 중성지방 함량은 20개월령 이후 급속히 증가됨을 보였고, 중성지방 함량의 증가와 함께 ACSS2, GPAT 및 DGAT 유전자의 발현이 증가됨을 확인하였다. 특히, 12개월령과 비교하여 ACSS2 유전자의 발현은 비육후기인 27개월령에서 7.3배, GPAT는 6.9배 증가되었다. 이러한 중성지방 함량의 증가와 중성지방 생합성 관련 유전자의 발현량 증가는 비육후기에 있어 고 에너지 사료 급여로 인한 지방산의 증가와, 증가된 지방산의 이용성 증대로 축적 형태인 중성지방 합성이 증가되는 것으로 판단되어 진다. 또한 중성지방 생합성 관련 유전자인 ACSS2, GPAT 및 DGAT 유전자의 발현증가는 근육 내 지방 축적과 밀접히 관련되어 있으며, 근내지방 축적과 관련되어 주요한 후보 유전자로 생각되어 진다.

• Journal of Plant Biotechnology
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• 제39권4호
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• pp.242-252
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• 2012

Seed storage oils are essential resources for not only human and animal diets but also industrial applications. The primary goal of this study was to increase seed oil content through comparative analysis of two seed-specific promoters, AtFAE1 from Arabidopsis Fatty Acid Elongase 1 gene and BnNapin from Brassica napus seed storage protein gene. AtWRI1 and AtDGAT1 genes encoding an AP2-type transcription factor and a Diacylglycerol Acyltransferase 1 enzyme, respectively, were expressed under the control of AtFAE1 and BnNapin promoters in Arabidopsis. The total seed oil content in all transgenic plants was increased by 8-11% compared with wild-type seeds. The increased level of oil content in AtWRI1 and AtDGAT1 transgenic lines under the control of both promoters was similar, although the activity of the BnNapin promoter is much stronger than that of AtFAE1 promoter in the mature stage of developing seeds where storage oil biosynthesis occurs at a maximum rate. This result demonstrates that the AtFAE1 promoter as well as the BnNapin promoter can be used to increase the seed oil content in transgenic plants.

• Journal of Microbiology and Biotechnology
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• 제18권11호
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• pp.1785-1788
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• 2008

Phenylpyropenes A, B, and C, isolated from Penicillium griseofulvum F1959, inhibited DGAT in rat liver microsomes with $IC_{50}$ values of $78.7{\pm}1.6$, $21.7{\pm}0.2$, and $11.04{\pm}0.2{\mu}M$, respectively. In addition, a kinetic analysis using a Lineweaver-Burk plot revealed that phenylpyropene C was a noncompetitive inhibitor of DGAT. The apparent Michaelis constant ($K_m$) value and inhibition constant ($K_i$) value were calculated to be $8{\mu}M$ and $10.48{\mu}M$, respectively. Moreover, phenylpyropene C inhibited triglyceride formation in HepG2 cells.

• Food Science and Biotechnology
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• 제18권1호
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• pp.95-102
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• 2009

The aerial parts of garland (Chrysanthemum coronarium L.) were extracted in 80% aqueous methanol (MeOH) and the concentrated extract was then partitioned using ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. EtOAc and n-BuOH fractions resulted in 4 glycerides with the application of octadecyl silica gel and silica gel column chromatography. The chemical structures of the glycerides were determined using several spectroscopic methods, including nuclear magnetic resonance (NMR) and mass spectrometry (MS) as (2S)-1-O-palmitoyl-sn-glycerol (1), (2S)-1-O-oleoyl-2-O-oleoyl- 3-O-$\beta$-D-galactopyranosyl-sn-glycerol (2), (2S)-1-O-palmitoyl-2-O-linoleoyl-3-O-phosphorouscholine-sn-glycerol (3), and (2S)-1-O-linolenoyl-2-O-palmitoyl-3-O-[$\alpha$-D-galactopyrasyl-($1{\rightarrow}6$)-$\beta$-D-galactopyranosyl]-sn-glycerol (4). The free fatty acids of these glycerides were determined with gas chromatography (GC)-MS analysis following alkaline hydrolysis and methylation. These glycerides demonstrated an inhibitory effect on acyl-CoA: cholesterol acyltransferase (ACAT, compound 1: $45.6{\pm}0.2%$ at $100{\mu}g/mL$), diacylglycerol acyltransferase (DGAT, compound 1: $59.1{\pm}0.1%$ at $25{\mu}g/mL$), farnesyl protein transferase (FPTase, compound 2: $98.0{\pm}0.1%$; compound 3: $55.2{\pm}0.1%$ at $100{\mu}g/mL$), and $\beta$-secretase ($IC_{50}$, compound 4: $2.6{\mu}g/mL$) activity. This paper is the first report on the isolation of these glycerides from garland and their inhibitory activity on ACAT, DGAT, FPTase, and $\beta$-secretase.

• BMB Reports
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• 제50권7호
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• pp.367-372
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• 2017

Monoacylglycerol acyltransferase 1 (MGAT) is a microsomal enzyme that catalyzes the synthesis of diacylglycerol (DAG) and triacylglycerol (TAG). However, the subcellular localization and catalytic function domain of this enzyme is poorly understood. In this report, we identified that murine MGAT1 localizes to the endoplasmic reticulum (ER) under normal conditions, whereas MGAT1 co-localize to the lipid droplets (LD) under conditions of enriching fatty acids, contributing to TAG synthesis and LD expansion. For the enzyme activity, both the N-terminal transmembrane domain and catalytic HPHG motif are required. We also show that the transmembrane domain of MGAT1 consists of two hydrophobic regions in the N-terminus, and the consensus sequence FLXLXXXn, a putative neutral lipid-binding domain, exists in the first transmembrane domain. Finally, MGAT1 interacts with DGAT2, which serves to synergistically increase the TAG biosynthesis and LD expansion, leading to enhancement of lipid accumulation in the liver and fat.