• Title/Summary/Keyword: metE gene

Search Result 33, Processing Time 0.025 seconds

Genetic Characterization of Molecular Targets in Korean Patients with Gastrointestinal Stromal Tumors

  • Park, Joonhong;Yoo, Han Mo;Sul, Hae Jung;Shin, Soyoung;Lee, Seung Woo;Kim, Jeong Goo
    • Journal of Gastric Cancer
    • /
    • v.20 no.1
    • /
    • pp.29-40
    • /
    • 2020
  • Purpose: Gastrointestinal stromal tumors (GISTs) frequently harbor activating gene mutations in either KIT or platelet-derived growth factor receptor A (PDGFRA) and are highly responsive to several selective tyrosine kinase inhibitors. In this study, a targeted next-generation sequencing (NGS) assay with an Oncomine Focus Assay (OFA) panel was used for the genetic characterization of molecular targets in 30 Korean patients with GIST. Materials and Methods: Using the OFA that enables rapid and simultaneous detection of hotspots, single nucleotide variants (SNVs), insertion and deletions (Indels), copy number variants (CNVs), and gene fusions across 52 genes relevant to solid tumors, targeted NGS was performed using genomic DNA extracted from formalin-fixed and paraffin-embedded samples of 30 GISTs. Results: Forty-three hotspot/other likely pathogenic variants (33 SNVs, 8 Indels, and 2 amplifications) in 16 genes were identified in 26 of the 30 GISTs. KIT variants were most frequent (44%, 19/43), followed by 6 variants in PIK3CA, 3 in PDGFRA, 2 each in JAK1 and EGFR, and 1 each in AKT1, ALK, CCND1, CTNNB1, FGFR3, FGFR4, GNA11, GNAQ, JAK3, MET, and SMO. Based on the mutation types, majority of the variants carried missense mutations (60%, 26/43), followed by 8 frameshifts, 6 nonsense, 1 stop-loss, and 2 amplifications. Conclusions: Our study confirmed the advantage of using targeted NGS with a cancer gene panel to efficiently identify mutations associated with GISTs. These findings may provide a molecular genetic basis for developing new drugs targeting these gene mutations for GIST therapy.

Arabidopsis AHL Gene Encodes a 3'(2'),5'-Bisphosphate Nucleotidase Sensitive to Toxic Heavy Metal Ions

  • Cheong, Jong-Joo;Kwon, Hawk-Bin
    • Journal of Applied Biological Chemistry
    • /
    • v.42 no.4
    • /
    • pp.169-174
    • /
    • 1999
  • Arabidopsis AHL gene contains 4 exons encoding a putative protein highly homologous to the yeast salt-sensitive enzyme HAL2, a 3'(2'),5'-bisphosphate nucleotidase involving in reductive sulfate assimilation. AHL cDNA complemented yeast met22 (hal2) mutant. AHL fusion protein expressed in E. coli exhibited $Mg^{2+}$-dependent, 3'-phosphoadenosine 5'-phosphate (PAP)-specific phosphatase activity. $Li^+,\;Na^+,\;K^+$ and $Ca^{2+}$ ions inhibit the enzyme activity by competing with $Mg^{2+}$ for the active site of the enzyme. The enzyme activity was also sensitive to ${\mu}M$ concentrations of toxic heavy metal ions such as $Cd^{2+},\;Cu^{2+}$ and $Zn^{2+}$, but was not recovered by addition of more $Mg^{2+}$ ions, suggesting that these ions inactivate the enzyme with a mechanism other than competition with $Mg^{2+}$ ions. Inhibition of the AHL enzyme activity may result in accumulation of PAP, which is highly toxic to the cell. Thus, the AHL enzyme could be one of the intial targets of heavy metal toxicity in plants.

  • PDF

Function of hepatocyte growth factor in gastric cancer proliferation and invasion

  • Koh, Sung Ae;Lee, Kyung Hee
    • Journal of Yeungnam Medical Science
    • /
    • v.37 no.2
    • /
    • pp.73-78
    • /
    • 2020
  • Cancer incidence has been increasing steadily and is the leading cause of mortality worldwide. Gastric cancer is still most common malignancy in Korea. Cancer initiation and progression are multistep processes involving various growth factors and their ligands. Among these growth factors, we have studied hepatocyte growth factor (HGF), which is associated with cell proliferation and invasion, leading to cancer and metastasis, especially in gastric cancer. We explored the intercellular communication between HGF and other surface membrane receptors in gastric cancer cell lines. Using complimentary deoxyribonucleic acid microarray technology, we found new genes associated with HGF in the stomach cancer cell lines, NUGC-3 and MKN-28, and identified their function within the HGF pathway. The HGF/N-methyl-N'-nitroso-guanidine human osteosarcoma transforming gene (c-MET) axis interacts with several molecules including E-cadherin, urokinase plasminogen activator, KiSS-1, Jun B, and lipocalin-2. This pathway may affect cell invasion and metastasis or cell apoptosis and is therefore associated with tumorigenesis and metastasis in gastric cancer.

Effects of Dietary Chromium Methionine on Growth Performance, Carcass Composition, Meat Colour and Expression of the Colour-related Gene Myoglobin of Growing-finishing Pigs

  • Li, Y.S.;Zhu, N.H.;Niu, P.P.;Shi, F.X.;Hughes, C.L.;Tian, G.X.;Huang, R.H.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.26 no.7
    • /
    • pp.1021-1029
    • /
    • 2013
  • To investigate the effect of dietary chromium (Cr) as Cr methionine (CrMet) on growth performance, carcass traits, pork quality, meat colour and expression of meat colour-related genes in growing-finishing pigs, 189 crossbred Duroc${\times}$(Landrace${\times}$Yorkshire) growing-finishing pigs (male, castrated, average initial BW $74.58{\pm}1.52$ kg) were selected and randomly allocated into four groups. Dietary treatments per kg of feed were as follows: 0 (CT), 0.3 mg/kg (T1), 0.6 mg/kg (T2) and 0.9 mg/kg (T3) Cr (in the form of CrMet; as-fed basis), and each treatment was replicated five times with 8 to 10 pigs per replicate pen. During the 28 d of the experiment, both the ADG and the ADFI increased linearly (p<0.05) as the level of dietary Cr increased. The F/G ratio decreased linearly (p<0.05). As dietary Cr increased, loin muscle areas (linear, p = 0.013) and average backfat thickness (linear, p = 0.072) decreased. Shear force (linear, p = 0.070) and Commission Internationale de I'$\acute{E}$clairage (CIE) redness (quadratic, p = 0.028) were increased. In addition, CIE Lightness (quadratic, p = 0.053) were decreased as dietary Cr increased. As dietary Cr increased, total myglobin (Mb) content (quadratic, p = 0.015) and the mb mRNA levels (quadratic, p = 0.046) in longissimus muscles of pigs were up-regulated. In conclusion, supplementation of dietary Cr improved growth and meat colour, but increased shear force and decreased IMF reduced palatability of longissimus muscles. Moreover, the increasing total Mb content and mb mRNA levels indicated that CrMet dietary supplementation may improve meat colour via up-regulating expression of the mb gene.

Purification and characterzation of the $\alpha$-L-Arabinofuranosidase from Escherichia coli Cells Harboring the Recombinant Plasmid pKMG11 (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus $\alpha$-L-Arabinofuranosidase의 정제 및 특성)

  • 엄수정;조쌍구;최용진
    • Microbiology and Biotechnology Letters
    • /
    • v.23 no.4
    • /
    • pp.446-453
    • /
    • 1995
  • $\alpha $-Arabinofuranosidase was produced by E. coli HB101 haboring the recombinant plasmid pKMG11 which contained the arfI gene of Bacillus stearothermophilus. The maximum production of the enzyme was observed when E. coli HB101 cells were grown at 37$\circ$C for 20 hours in the medium containing 0.5% arabinose, 1.0% tryptone, 0.5% yeast extract, and 1% NaCl. The $\ALPHA $-arabinofuranosidase produced was purified to homogeneity using a combination of 20-50% ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange column chromatography and Sepharose 6B-100 gel filtration. The purified enzyme was most active at 55$\circ$C and pH 6.5. The K$_{m}$ and V$_{max}$ values of the enzyme on $\rho $-nitrophenyl-$\alpha $-arabinofuranoside was determined to be 2.99 mM and 0.43 $\mu $mole/min (319.74 $\mu $mole/min/mg), respectively. The pI value was 4.5. The molecular weight of the native protein was estimated to be 289 kDa. The SDS-polyacrylamide gel clectrophoresis analysis suggested that the functional protein was a trimer of the 108 kDa identical subunits. The N-terminal amino acid sequence of the a-arabinofuranosidase was identified as X-Ser-Thr-Ala-Pro-Arg( \ulcorner )-Ala-Thr-Met-Val-Ile-Asp-X-Ala-Phe.

  • PDF

ermK Leader Peptide : Amino Acid Sequence Critical for Induction by Erythromycin

  • Kwon, Ae-Ran;Min, Yu-Hong;Yoon, Eun-Jeong;Kim, Jung-A;Shim, Mi-Ja;Choi, Eung-Chil
    • Archives of Pharmacal Research
    • /
    • v.29 no.12
    • /
    • pp.1154-1157
    • /
    • 2006
  • The ermK gene from Bacillus lichenformis encodes an inducible rRNA methylase that confers resistance to the macrolide-lincosamide-streptogramin B antibiotics. The ermK mRNA leader sequence has a total length of 357 nucleotides and encodes a 14-amino acid leader peptide together with its ribosome binding site. The secondary structure of ermK leader mRNA and a leader peptide sequence have been reported as the elements that control expression. In this study, the contribution of specific leader peptide amino acid residues to induction of ermK was studied using the PCR-based megaprimer mutation method. ermK methylases with altered leader peptide codons were translationally fused to E. coli ${\beta}-galactosidase$ reporter gene. The deletion of the codons for Thr-2 through Ser-4 reduced inducibility by erythromycin, whereas that for Thr-2 and His-3 was not. The replacement of the individual codons for Ser-4, Met-5 and Arg-6 with termination codon led to loss of inducibility, but stop mutation of codon Phe-9 restored inducibility by erythromycin. Collectively, these findings suggest that the codons for residue 4, 5 and 6 comprise the critical region for induction. The stop mutation at Leu-7 expressed constitutively ermK gene. Thus, ribosome stalling at codon 7 appears to be important for ermK induction.

Cloning, Sequencing and Characterization of Mitochondrial Control Region of the Domestic Silkwom, Bombyx mori

  • Lee, Jin-Sung;Kim, Ki-Hwan;Hoe, Hyang-Sook;Park, Jae-Heung;Kang, Seok-Woo;Lee, Sang-Han;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.2 no.1
    • /
    • pp.87-89
    • /
    • 2001
  • The nucleotide sequence of the domestic silkworm (Bombyx mori) mitochondrial (mt) control region and its flanking genes was determined from PCR clones. The control region of the silkworm mt genome was located between the small ribosomal RNA gene and transfer RN $A^{Met}$. This 499 bp control region hale 95.4% A+T content. Extensive comparative analysis studies performed with similar control region of other insect genomes could not reveal a highly conserved region containing conserved motifs of animal mito-chondrial genome. The remarkable feature that found in this control region was the presence of tandem motifs containing nine repetitive sequences. The potential usefulness of this motif sequences for Bombyx species or their taxonomically related species is enhanced by its unique localization in the maternally inheritance mitochondrial molecule.e.

  • PDF

MicroRNA-214 Regulates the Acquired Resistance to Gefitinib via the PTEN/AKT Pathway in EGFR-mutant Cell Lines

  • Wang, Yong-Sheng;Wang, Yin-Hua;Xia, Hong-Ping;Zhou, Song-Wen;Schmid-Bindert, Gerald;Zhou, Cai-Cun
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.1
    • /
    • pp.255-260
    • /
    • 2012
  • Patients with non-small cell lung cancer (NSCLC) who have activating epidermal growth factor receptor (EGFR) mutations derive clinical benefit from treatment with EGFR-tyrosine kinase inhibitors ((EGFR-TKIs)-namely gefitinib and erlotinib. However, these patients eventually develop resistance to EGFR-TKIs. Despite the fact that this acquired resistance may be the result of a secondary mutation in the EGFR gene, such as T790M or amplification of the MET proto-oncogene, there are other mechanisms which need to be explored. MicroRNAs (miRs) are a class of small non-coding RNAs that play pivotal roles in tumorigenesis, tumor progression and chemo-resistance. In this study, we firstly successfully established a gefitinib resistant cell line-HCC827/GR, by exposing normal HCC827 cells (an NSCLC cell line with a 746E-750A in-frame deletion of EGFR gene) to increasing concentrations of gefitinib. Then, we found that miR-214 was significantly up-regulated in HCC827/GR. We also showed that miR-214 and PTEN were inversely expressed in HCC827/GR. Knockdown of miR-214 altered the expression of PTEN and p-AKT and re-sensitized HCC827/GR to gefitinib. Taken together, miR-214 may regulate the acquired resistance to gefitinib in HCC827 via PTEN/AKT signaling pathway. Suppression of miR-214 may thus reverse the acquired resistance to EGFR-TKIs therapy.

High Density Lipoprotein from Egg Yolk (EYHDL) Improves Dyslipidemia by Mediating Fatty Acids Metabolism in High Fat Diet-induced Obese Mice

  • Yu, Zhihui;Mao, Changyi;Fu, Xing;Ma, Meihu
    • Food Science of Animal Resources
    • /
    • v.39 no.2
    • /
    • pp.179-196
    • /
    • 2019
  • We investigated the effect of high density lipoprotein from egg yolk (EYHDL) on serum, hepatic and fecal lipid and fatty acids (FAs) levels and on gene expression involved in FAs metabolism. Male KM mice were fed either normal diet (ND; n=20), high fat diet (HFD; n=20), or high fat diet containing EYHDL (EYHDL; 0.6 mg/g, every day by oral gavage, n=20) for 100 days. At the end of the experiment, the effects of treatments on biochemical parameters, FAs profiles and involved gene expression were analyzed. Our results revealed that EYHDL markedly suppressed the body weight gain, accumulation of abdominal fat tissues, serum concentrations of LDL-cholesterol (LDL-C) and triglycerides, hepatic triglycerides and cholesterol accumulation, while increased serum concentration of HDL-cholesterol (HDL-C). EYHDL intake also increased total cholesterol (TC) excretions compared with HFD group. Moreover, it alleviated the severity of fatty liver and improved glucose and insulin tolerance compared with HFD. More importantly, EYHDL partially normalized FAs profiles in serum, liver and fecaces and neutralized the HFD-induced upregulation of SREBP-1c, Acaca, Fasn, GPAT and Scd1. In conclusion, our findings indicate that EYHDL may have the potential to improve metabolic disturbances that occur in HFD mice and can be considered as an appropriate dietary recommendation for the treatment of metabolic syndrome (MetS).

Applicable Feasibility of Fenton Oxidation and Zeolite Ion Exchange Processes for Removal of Non-Biodegradable Matters and Ammonia in Livestock Wastewater (축산폐수 중의 난분해성 물질 및 암모니아 제거를 위한 Fenton 산화와 Zeolite 이온교환 공정의 적용 가능성)

  • Cho, Chang-Woo;Kim, Byoung-Young;Chae, Soo-Choen;Chung, Paul-Gene
    • Journal of Korean Society on Water Environment
    • /
    • v.23 no.5
    • /
    • pp.600-606
    • /
    • 2007
  • Livestock wastewater containing concentrated organic matters and nutrients has been known as one of the major pollutants. It is difficult to apply the conventional activated sludge process to treat livestock wastewater because of high Non-biodegradable (NBD) matter and ammonia. The objectives of this study are to remove NBD matters including aromatic compounds and ammonia in livestock wastewater using Coagulation-Fenton oxidation-Zeolite (CFZ) processes and ascertain applicable feasibility in the field through pilot plant experiment. NBD matters and color remained in the treated water were removed over 92% by Fenton oxidation as the second treatment process. Ammonia was removed by over 99.5% in the zeolite ion exchange process as the last treatment method. From $UV_{254}$, $E_2/E_3$ ratio and GC/MS analyses of treated water at each process, the aromatic compound was converted to aliphatic and aromaticity was decreased. In pilot scale test, organics and ammonia removal efficiencies were not much different from the result of lab-scale test at various operation conditions. Furthermore, reaction time and dosage of Fenton reagent in pilot scale experiment reduced by 40 min and 50% rather than in lab-scale test. $BOD_5$, $COD_{Mn}$, SS, T-N and T-P of treated water in the pilot-scale experiment also met the effluent standards.