DOCSLIB.ORG

Characterization and Mapping of the Bovine FBP1 Gene

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization and Mapping of the Bovine FBP1 Gene 1319 Asian-Aust. J. Anim. Sci. Vol. 20, No. 9 : 1319 - 1326 September 2007 www.ajas.info Characterization and Mapping of the Bovine FBP1 Gene H. Guo1, 2, W-S. Liu3, A. Takasuga4, K. Eyer3, E. Landrito3, S-Z. Xu1, *, X. Gao1 and H-Y. Ren1 1 Laboratory of Molecular Biology and Bovine Breeding, Institute Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100094, P. R. China ABSTRACT :Fructose-1,6-bisphosphatase (FBP1) is a key regulatory enzyme of gluconeogenesis that catalyzes the hydrolysis of fructose-1,6-bisphosphate to generate fructose-6-phosphate and inorganic phosphate. Deficiency of fructose-1, 6-bisphosphatase is associated with fasting hypoglycemia and metabolic acidosis. The enzyme has been shown to occur in bacteria, fungi, plants and animals. The bovine FBP1 gene was cloned and characterized in this study. The full length (1,241 bp) FBP1 mRNA contained an open reading frame (ORF) encoding a protein of 338 amino acids, a 63 bp 5’ untranslated region (UTR) and a 131 bp 3’ UTR. The bovine FBP1 gene was 89%, 85%, 82%, 82% and 74% identical to the orthologs of pig, human, mouse, rat and zebra fish at mRNA level, and 97%, 96%, 94%, 93% and 91% identical at the protein level, respectively. This gene was broadly expressed in cattle with the highest level in testis, and the lowest level in heart. An intronic single nucleotide polymorphism (SNP) (A/G) was identified in the 5th intron of the bovine FBP1 gene. Genotyping of 133 animals from four beef breeds revealed that the average frequency for allele A (A-base) was 0.7897 (0.7069-0.9107), while 0.2103 (0.0893-0.2931) for allele B (G-base). Our preliminary association study indicated that this SNP is significantly associated with traits of Average Daily Feed Intake (ADFI) and Carcass Length (CL) (p<0.01). In addition, the FBP1 gene was assigned on BTA8 by a hybrid radiation (RH) mapping method. (Key Words : Different Expression, Localization, FBP1, mRNA, Cattle) INTRODUCTION leading to impaired liver gluconeogenesis, have been identified and in several cases the specific mutations Fructose-1,6-bisphosphatase (EC 3.1.3.11) (FBPase) is a causing the disease could be traced (Kikawa et al., 1994; key regulatory enzyme of gluconeogenesis that catalyzes Herzog et al., 1999). Besides the human FBP1 gene the hydrolysis of fructose-1,6-bisphosphate to generate (Solomon et al., 1988, El-Maghrabi et al., 1993, 1995), the fructose-6-phosphate and inorganic phosphate. The enzyme gene orthologs have been characterized in several other has been shown to occur in a variety of species, including species, including mouse (Stein et al., 2001), pig (Marcus et bacteria, fungi, plants and animals (Tillmann et al., 2000; al., 1982), rat (El-Maghrabi et al., 1988, 1991), and zebra Stein et al., 2001). There are two genes, FBP1 (liver fish (Woods et al., 2005). The human, mouse and pig FBP1 FBPase) and FBP2 (muscle FBPase) that code for FBPases gene encode a polypeptide of 338 amino acids (aa) (Marcus in mammals. FBP1 is expressed predominantly in liver, et al., 1982; Skalecki et al., 1999; Stein et al., 2001), kidney, lung and monocytes (El-Maghrabi et al., 1991, whereas zebra fish and rat (El-Maghrabi et al., 1991) FBP1 1995), while FBP2 is expressed predominantly in muscle genes encode polypeptides of 337 aa and 363 aa, (Tillmann et al., 2000). Patients with defective FBP1, respectively. In humans, a report found that a hereditary lack of liver FBPase leads to FBPase deficiency, which is * Corresponding Author: Shang-zhong Xu. Tel: +86-10-62816065, an autosomal recessive condition associated with Fax: +86-10-62817806, E-mail: [email protected] hypoglycemia and metabolic acidosis (Stein et al., 2001). 2 Department of Animal Science, Tianjin Agricultural College, Molecular genetic analyses have revealed several mutations TianJin 300384, P. R. China. 3 Department of Animal Biotechnology, University of from the coding region in FBP1 as possible causes for the Agriculture, Biotechnology and Natural Resources, University of problem of hypoglycemia and metabolic acidosis (Kikawa Nevada, Reno, Nevada 89557, USA. et al., 1997; Herzog et al., 1999). In bovine industry, the 4 Shirakawa Institute of Animal Genetics Odakura, Nishigo, metabolic acidosis and hypoglycemia (such as acetonemia) Nishi-shirakawa, Fukushima 961-8061, Japan. happens frequently and is usually resolved by dietary means Received December 4, 2006; Accepted April 26, 2007 1320 Guo et al. (2007) Asian-Aust. J. Anim. Sci. 20(9):1319-1326 Table 1. Primers used for the bovine FBP1 and GAPDH genes Gene name Primer names Primer sequences Tm (°C) Size (bp) Binding region FBP11 F 5'-AGGCTATGCGAAGGACTTTG-3' 62 1,157 exon5 R 5'-ATAGACTAGGGTGCGGTGTA-3' exon6 F1 5'-TCATCTCTCCTTCCACCTCT-3' 59 507 intron5 3’-RACE 5'-GCCCAGCCCACACCAAATGGAATTG-3' 68 154 3’-UTR 5’-RACE 5'-GTTGAGCAGTTGGGTCATCTCGCC-3' 68 181 exon1 F2 5'-CGACACCAATATCGTCACCG-3' 60 869 exon1 R2 5'-TAGCCATTCCTCCTGCCTTC-3' exon7 Real-time PL 5'-ATGGCAGTGCCACTATGTTG-3' 60 156 exon4 Real-time PR 5'-AGTCCTTCGCATAGCCTTCA-3' exon5 Taqman probe 5'-CATGGCGAATGGAGTCAACTGCTT-3' exon4 GAPDH Real-time PL 5'-GATCAAGAAGGTGGTGAAGC-3' 60 156 exon10 Real-time PR 5'-TTGACAAAGTGGTCGTTGAG-3' exon11 Taqman probe 5'-CCTCTCAAGGGCATTCTAGGCTACA-3' exon10 1 The primer pair F-R was used to isolate the 1,157 bp length genomic DNA fragment of the FBP1, the primer pair F1-R was used for RH mapping. The primer pair F2-R2 for isolating CDS. Real-time PL and Real-time PR were used for the gene expression patterns. (Schonewille et al., 1999; McNeill et al., 2002). populations were used to analyze the FBP1 allelic Understanding the molecular mechanism of acidosis and the frequencies. genes involved in hypocalcaemia will provide the molecular Approximately eleven different tissues (heart, liver, lung, basis for finding new solutions to this problem. In this paper, spleen, intestine, kidney, loin-muscle, rumen, testis, lymph, we report the full-length cDNA sequence of the bovine and thymus) were collected from mature Simmental cattle FBP1 gene, its predicted aa sequence, genomic structure, (Inner Mongolia Province, China) at slaughter. expression patterns and single nucleotide polymorphisms (SNPs) as an initial step for identifying the relationship Statistical analyses between FBP1 and the acidosis-related performance in The following traits, Backfat Thickness (BF), Carcass cattle. Weight (CW), Meat Percent (MP), Average Daily Gain (ADG), Average Daily Feed Intake (ADFI), Ratio of Feed- MATERIALS AND METHODS to-Meat (RFM), Beginning Average Daily Gain (BADG), Finishing Average Daily Gain (FADG), Loin-Muscle Area Source of animals and tissues (LMA) and Carcass Length (CL) were measured or A total of 51 crossbred steers (Simmental crossbred calculated. The collected data were analyzed using a GLM with indigenous female yellow cattle in China) (405±50.5 procedure of SAS (SAS Institute Inc. Cary, NC, USA) kg; 30±2 months of age) were selected randomly and according to the following model: housed in a concrete-floored cowshed (in a single pan for each animal). The pre-trial period (for adaptation to Yj =卩+G i+bijwij+Sij treatment) was 15 days, and the test period was 180 days. Steers were fed according to the NY/T 815-2004 feeding Where Yj stands for observed value;卩，population standard of beef cattle (Agricultural Department of People's mean; Gi , ith genotype; bij: regression coefficient; wij: Republic of China, 2004), and had free access to water and weight, for growth traits the wij stands for starting weight, feed during the entire 195 days of the experimental period. for meat traits, slaughter weight.;司，random error. All animals were weighed on days 1, 75 and 195 at 8:00 AM, with prior removal of feed and water (12 h). The DNA extraction and RNA extraction remainders of feed each day were recorded at 10:00 PM Blood samples were collected. The routine phenol each day. Immediately after the trial; steers were harvested chloroform extraction method was used to isolate the at a processing facility (KeErQing Beef cattle Co., Ltd. P. R. genomic DNA that was diluted to 50 ng/卩l for PCR. Total China). Carcass traits were measured according to the RNA was extracted from eleven tissues using the TRIzol criterion GB/T 17238-1998 Cutting Standard of fresh and reagent kit (Life Technologies, Grand Island, NE, USA). chilled beef in China (China Standard Publishing House). A total of 51 crossbred steers were used to investigate Primer design associations of gene variant with traits. In addition, a total The mRNA sequence for the human FBP1 of 82 animals including Simmental (n = 28), Angus (n = 25), (NM_000507) gene was obtained from NCBI, and was Hereford (n = 29) randomly selected from commercial Guo et 시. (2007) Asian-Aust. J. Anim. Sci. 20(9):1319-1326 1321 Table 2. Allele frequencies of the bovine FBP1 gene in different 60°C (for annealing and polymerization). Relative mRNA breeds quantification was performed using standard curves Allele frequency Breeds No. of animals generated with heart tissue total RNA. PCRs were A B performed in triplicate and gene expression levels were Simmental 28 0.91 0.09 quantified relative to the expression of GAPDH using Gene Angus 25 0.76 0.24 Hereford 29 0.71 0.29 Expression Macro software (Bio-Rad) employing the Cross simmental 51 0.78 0.22 comparative C (△△Ct) value method. Expression levels were considered not detectable when the C value of the employed to search for bovine expressed sequence tags targeted gene exceeded 35 in the sample tissue. (ESTs) in the EST-others database through standard BLAST (http://www.ncbi.nlm.nih.gov/blast/) algorithm. The bovine Isolation and sequencing of the gene fragment ESTs that have more than 80% similarity with the The amplified products were obtained for FBP1 in a 20 corresponding human mRNA were candidates, assembled il reaction volume consisting of 50 ng of bovine genomic into a contig and used to design the gene-specific primers.
Load more

Recommended publications
  • The Switch from Fermentation to Respiration in Saccharomyces Cerevisiae Is Regulated by the Ert1 Transcriptional Activator/Repressor
    INVESTIGATION The Switch from Fermentation to Respiration in Saccharomyces cerevisiae Is Regulated by the Ert1 Transcriptional Activator/Repressor Najla Gasmi,* Pierre-Etienne Jacques,† Natalia Klimova,† Xiao Guo,§ Alessandra Ricciardi,§ François Robert,†,** and Bernard Turcotte*,‡,§,1 ‡Department of Medicine, *Department of Biochemistry, and §Department of Microbiology and Immunology, McGill University Health Centre, McGill University, Montreal, QC, Canada H3A 1A1, †Institut de recherches cliniques de Montréal, Montréal, QC, Canada H2W 1R7, and **Département de Médecine, Faculté de Médecine, Université de Montréal, QC, Canada H3C 3J7 ABSTRACT In the yeast Saccharomyces cerevisiae, fermentation is the major pathway for energy production, even under aerobic conditions. However, when glucose becomes scarce, ethanol produced during fermentation is used as a carbon source, requiring a shift to respiration. This adaptation results in massive reprogramming of gene expression. Increased expression of genes for gluconeogenesis and the glyoxylate cycle is observed upon a shift to ethanol and, conversely, expression of some fermentation genes is reduced. The zinc cluster proteins Cat8, Sip4, and Rds2, as well as Adr1, have been shown to mediate this reprogramming of gene expression. In this study, we have characterized the gene YBR239C encoding a putative zinc cluster protein and it was named ERT1 (ethanol regulated transcription factor 1). ChIP-chip analysis showed that Ert1 binds to a limited number of targets in the presence of glucose. The strongest enrichment was observed at the promoter of PCK1 encoding an important gluconeogenic enzyme. With ethanol as the carbon source, enrichment was observed with many additional genes involved in gluconeogenesis and mitochondrial function. Use of lacZ reporters and quantitative RT-PCR analyses demonstrated that Ert1 regulates expression of its target genes in a manner that is highly redundant with other regulators of gluconeogenesis.
    [Show full text]
  • Analysis of Gene Expression Data for Gene Ontology
    ANALYSIS OF GENE EXPRESSION DATA FOR GENE ONTOLOGY BASED PROTEIN FUNCTION PREDICTION A Thesis Presented to The Graduate Faculty of The University of Akron In Partial Fulfillment of the Requirements for the Degree Master of Science Robert Daniel Macholan May 2011 ANALYSIS OF GENE EXPRESSION DATA FOR GENE ONTOLOGY BASED PROTEIN FUNCTION PREDICTION Robert Daniel Macholan Thesis Approved: Accepted: _______________________________ _______________________________ Advisor Department Chair Dr. Zhong-Hui Duan Dr. Chien-Chung Chan _______________________________ _______________________________ Committee Member Dean of the College Dr. Chien-Chung Chan Dr. Chand K. Midha _______________________________ _______________________________ Committee Member Dean of the Graduate School Dr. Yingcai Xiao Dr. George R. Newkome _______________________________ Date ii ABSTRACT A tremendous increase in genomic data has encouraged biologists to turn to bioinformatics in order to assist in its interpretation and processing. One of the present challenges that need to be overcome in order to understand this data more completely is the development of a reliable method to accurately predict the function of a protein from its genomic information. This study focuses on developing an effective algorithm for protein function prediction. The algorithm is based on proteins that have similar expression patterns. The similarity of the expression data is determined using a novel measure, the slope matrix. The slope matrix introduces a normalized method for the comparison of expression levels throughout a proteome. The algorithm is tested using real microarray gene expression data. Their functions are characterized using gene ontology annotations. The results of the case study indicate the protein function prediction algorithm developed is comparable to the prediction algorithms that are based on the annotations of homologous proteins.
    [Show full text]
  • Role of RUNX1 in Aberrant Retinal Angiogenesis Jonathan D
    Page 1 of 25 Diabetes Identification of RUNX1 as a mediator of aberrant retinal angiogenesis Short Title: Role of RUNX1 in aberrant retinal angiogenesis Jonathan D. Lam,†1 Daniel J. Oh,†1 Lindsay L. Wong,1 Dhanesh Amarnani,1 Cindy Park- Windhol,1 Angie V. Sanchez,1 Jonathan Cardona-Velez,1,2 Declan McGuone,3 Anat O. Stemmer- Rachamimov,3 Dean Eliott,4 Diane R. Bielenberg,5 Tave van Zyl,4 Lishuang Shen,1 Xiaowu Gai,6 Patricia A. D’Amore*,1,7 Leo A. Kim*,1,4 Joseph F. Arboleda-Velasquez*1 Author affiliations: 1Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, 20 Staniford St., Boston, MA 02114 2Universidad Pontificia Bolivariana, Medellin, Colombia, #68- a, Cq. 1 #68305, Medellín, Antioquia, Colombia 3C.S. Kubik Laboratory for Neuropathology, Massachusetts General Hospital, 55 Fruit St., Boston, MA 02114 4Retina Service, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, 243 Charles St., Boston, MA 02114 5Vascular Biology Program, Boston Children’s Hospital, Department of Surgery, Harvard Medical School, 300 Longwood Ave., Boston, MA 02115 6Center for Personalized Medicine, Children’s Hospital Los Angeles, Los Angeles, 4650 Sunset Blvd, Los Angeles, CA 90027, USA 7Department of Pathology, Harvard Medical School, 25 Shattuck St., Boston, MA 02115 Corresponding authors: Joseph F. Arboleda-Velasquez: [email protected] Ph: (617) 912-2517 Leo Kim: [email protected] Ph: (617) 912-2562 Patricia D’Amore: [email protected] Ph: (617) 912-2559 Fax: (617) 912-0128 20 Staniford St. Boston MA, 02114 † These authors contributed equally to this manuscript Word Count: 1905 Tables and Figures: 4 Diabetes Publish Ahead of Print, published online April 11, 2017 Diabetes Page 2 of 25 Abstract Proliferative diabetic retinopathy (PDR) is a common cause of blindness in the developed world’s working adult population, and affects those with type 1 and type 2 diabetes mellitus.
    [Show full text]
  • Clinical and Molecular Characterization of Patients with Fructose 1,6-Bisphosphatase Deficiency
    Article Clinical and Molecular Characterization of Patients with Fructose 1,6-Bisphosphatase Deficiency Niu Li 1,†, Guoying Chang 2,†, Yufei Xu 1, Yu Ding 2, Guoqiang Li 1, Tingting Yu 1, Yanrong Qing 1, Juan Li 2, Yiping Shen 1,3, Jian Wang 1,* and Xiumin Wang 2,* 1 Department of Medical Genetics and Molecular Diagnostic Laboratory, Shanghai Children’s Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China; [email protected] (N.L.); [email protected] (Y.X.); [email protected] (G.L.); [email protected] (T.Y.); [email protected] (Y.Q.); [email protected] (Y.S.) 2 Department of Endocrinology and Metabolism, Shanghai Children’s Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China; [email protected] (G.C.); [email protected] (Y.D.); [email protected] (J.L.) 3 Department of Laboratory Medicine, Boston Children’s Hospital, Boston, MA 02115, USA * Correspondence: [email protected] (J.W.); [email protected] (X.W.); Tel.: +86-21-3862-6161 (J.W. & X.W.); Fax: +86-21-5875-6923 (J.W. & X.W.) † These authors contributed equally to this work. Academic Editor: William Chi-shing Cho Received: 26 February 2017; Accepted: 17 April 2017; Published: 18 April 2017 Abstract: Fructose-1,6-bisphosphatase (FBPase) deficiency is a rare, autosomal recessive inherited disease caused by the mutation of the FBP1 gene, the incidence is estimated to be between 1/350,000 and 1/900,000. The symptoms of affected individuals are non-specific and are easily confused with other metabolic disorders.
    [Show full text]
  • Product Description SALSA® MLPA® Probemix P255-B1 ALDOB-FBP1 to Be Used with the MLPA General Protocol
    Product description version B1-01; Issued 14 January 2021 Product Description SALSA® MLPA® Probemix P255-B1 ALDOB-FBP1 To be used with the MLPA General Protocol. Version B1 No Changes. Catalogue numbers: P255-025R: SALSA MLPA Probemix P255 ALDOB-FBP1, 25 reactions. P255-050R: SALSA MLPA Probemix P255 ALDOB-FBP1, 50 reactions. P255-100R: SALSA MLPA Probemix P255 ALDOB-FBP1, 100 reactions. To be used in combination with a SALSA MLPA reagent kit and Coffalyser.Net data analysis software. MLPA reagent kits are either provided with FAM or Cy5.0 dye-labelled PCR primer, suitable for Applied Biosystems and Beckman/SCIEX capillary sequencers, respectively (see www.mrcholland.com). Certificate of Analysis Information regarding storage conditions, quality tests, and a sample electropherogram from the current sales lot is available at www.mrcholland.com. Precautions and warnings For professional use only. Always consult the most recent product description AND the MLPA General Protocol before use: www.mrcholland.com. It is the responsibility of the user to be aware of the latest scientific knowledge of the application before drawing any conclusions from findings generated with this product. General information The SALSA MLPA Probemix P255-B1 ALDOB-FBP1 is a research use only (RUO) assay for the detection of deletions or duplications in the ALDOB and FBP1, which are associated with Hereditary fructose intolerance. Hereditary fructose intolerance (HFI) is a rare recessive inherited disorder of carbohydrate metabolism, caused by catalytic deficiency of the aldolase B enzyme (ALDOB). The ALDOB enzyme plays a key role in glycolysis and gluconeogenesis and, in mammals, is preferentially expressed in the liver.
    [Show full text]
  • Datasheet: 0200-0058 Product Details
    Datasheet: 0200-0058 Description: MOUSE ANTI FOLATE BINDING PROTEIN Specificity: FOLATE BINDING PROTEIN Other names: FBP Format: Purified Product Type: Monoclonal Antibody Clone: SF75 (BGN/1210/75) Isotype: IgG2b Quantity: 0.2 mg Product Details Applications This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit www.bio-rad-antibodies.com/protocols. Yes No Not Determined Suggested Dilution ELISA Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using the appropriate negative/positive controls. Target Species Bovine Product Form Purified IgG - liquid Preparation Prepared by Protein A chromatography Buffer Solution Phosphate buffered saline Preservative 0.09% Sodium Azide (NaN ) Stabilisers 3 Approx. Protein IgG concentration 1.0mg/ml Concentrations Immunogen Purified Folate Binding Protein from bovine milk. External Database UniProt: Links P02702 Related reagents Entrez Gene: 432398 FBP1 Related reagents Page 1 of 3 Specificity Mouse anti Bovine folate binding protein antibody, clone SF75 recognizes folate binding protein isolated from bovine milk. Cross reactivity with folate binding proteins from other sources has not been determined. Soluble folate binding protein from milk serves to accumulate and stabilize reduced folate compounds in the milk and aid in the absorbtion of folates by the intestinal mucosa of neonates.
    [Show full text]
  • Far Upstream Element Binding Protein 1: a Commander of Transcription, Translation and Beyond
    Oncogene (2013) 32, 2907–2916 & 2013 Macmillan Publishers Limited All rights reserved 0950-9232/13 www.nature.com/onc REVIEW Far upstream element binding protein 1: a commander of transcription, translation and beyond J Zhang and QM Chen The far upstream binding protein 1 (FBP1) was first identified as a DNA-binding protein that regulates c-Myc gene transcription through binding to the far upstream element (FUSE) in the promoter region 1.5 kb upstream of the transcription start site. FBP1 collaborates with TFIIH and additional transcription factors for optimal transcription of the c-Myc gene. In recent years, mounting evidence suggests that FBP1 acts as an RNA-binding protein and regulates mRNA translation or stability of genes, such as GAP43, p27 Kip and nucleophosmin. During retroviral infection, FBP1 binds to and mediates replication of RNA from Hepatitis C and Enterovirus 71. As a nuclear protein, FBP1 may translocate to the cytoplasm in apoptotic cells. The interaction of FBP1 with p38/ JTV-1 results in FBP1 ubiquitination and degradation by the proteasomes. Transcriptional and post-transcriptional regulations by FBP1 contribute to cell proliferation, migration or cell death. FBP1 association with carcinogenesis has been reported in c-Myc dependent or independent manner. This review summarizes biochemical features of FBP1, its mechanism of action, FBP family members and the involvement of FBP1 in carcinogenesis. Oncogene (2013) 32, 2907–2916; doi:10.1038/onc.2012.350; published online 27 August 2012 Keywords: FBP1; FUSE; cancer INTRODUCTION IDENTIFICATION OF FBP1 The far upstream element binding protein 1 (FBP1) was first FBP1 was first cloned from a cDNA library generated from the discovered as a transcriptional regulator of the proto-oncogene undifferentiated human monoblastic line U937.6 Such an c-Myc.
    [Show full text]
  • A MAFG-Lncrna Axis Links Systemic Nutrient Abundance to Hepatic Glucose Metabolism
    ARTICLE https://doi.org/10.1038/s41467-020-14323-y OPEN A MAFG-lncRNA axis links systemic nutrient abundance to hepatic glucose metabolism Marta Pradas-Juni et al.# Obesity and type 2 diabetes mellitus are global emergencies and long noncoding RNAs (lncRNAs) are regulatory transcripts with elusive functions in metabolism. Here we show that a high fraction of lncRNAs, but not protein-coding mRNAs, are repressed during diet-induced 1234567890():,; obesity (DIO) and refeeding, whilst nutrient deprivation induced lncRNAs in mouse liver. Similarly, lncRNAs are lost in diabetic humans. LncRNA promoter analyses, global cistrome and gain-of-function analyses confirm that increased MAFG signaling during DIO curbs lncRNA expression. Silencing Mafg in mouse hepatocytes and obese mice elicits a fasting-like gene expression profile, improves glucose metabolism, de-represses lncRNAs and impairs mammalian target of rapamycin (mTOR) activation. We find that obesity-repressed LincIRS2 is controlled by MAFG and observe that genetic and RNAi-mediated LincIRS2 loss causes elevated blood glucose, insulin resistance and aberrant glucose output in lean mice. Taken together, we identify a MAFG-lncRNA axis controlling hepatic glucose metabolism in health and metabolic disease. #A full list of authors and their affiliations appears at the end of the paper. NATURE COMMUNICATIONS | (2020) 11:644 | https://doi.org/10.1038/s41467-020-14323-y | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | https://doi.org/10.1038/s41467-020-14323-y ellular and organism-level energy homeostasis and nutri- by MAFG and CRISPR–Cas9-mediated knockout, or antisense- Cent partitioning are instrumental for survival. In higher mediated RNA interference of LincIRS2 causes hyperglycemia, organisms, multi-organ systems evolved to react to shifts insulin resistance, likely caused by alterations in glucogenic in energy supply by storing (anabolic) or metabolizing (catabolic) gene expression in lean mice.
    [Show full text]
  • Long Noncoding RNA DANCR Regulates Proliferation and Migration by Epigenetically Silencing FBP1 in Tumorigenesis of Cholangiocar
    Wang et al. Cell Death and Disease (2019) 10:585 https://doi.org/10.1038/s41419-019-1810-z Cell Death & Disease ARTICLE Open Access Long noncoding RNA DANCR regulates proliferation and migration by epigenetically silencing FBP1 in tumorigenesis of cholangiocarcinoma Ni Wang1, Chongguo Zhang2, Wulin Wang3,JieLiu4,YangYu1,YouLi5, Mingjiong Zhang6,XianxiuGe1, Quanpeng Li1 and Lin Miao 1 Abstract Recently, long noncoding RNAs (lncRNAs) have been shown to play significant regulatory roles in human tumorigenesis. However, the biological function of lncRNAs in cholangiocarcinoma (CCA) remains largely unknown. In this study, DANCR was shown to be significantly upregulated in CCA. DANCR regulated the proliferation and migration of CCA cells in vitro. Moreover, downregulation of DANCR suppressed CCA cells proliferation in vivo. RNA-seq revealed that DANCR knockdown preferentially affected genes linked with cell proliferation and cell differentiation. Furthermore, mechanistic investigation validated that DANCR could bind EZH2 and modulate the histone methylation of promoter of FBP1, thereby regulating CCA cells growth and migration. Taken together, these results demonstrated the significant roles of DANCR in CCA and may provide a theoretical basis for clinical diagnosis and treatment of CCA. 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Introduction Long noncoding RNAs (lncRNAs), the RNA transcripts Cholangiocarcinoma (CCA), an extremely malignant longer than 200 nucleotides with little or no protein- tumor that arises from cholangiocytes, has been a major coding potential4,5, have been shown to exhibit critical health burden worldwide for decades1. Given the lack of function in tumorigenesis, including CCA6,7. In addition, sensitive indicators, the diagnosis of the majority of CCA as a regulator in the process of epigenetics, lncRNAs cases typically occurs at a late stage, resulting that patients could regulate gene expression in chromatin modification, with unresectable tumors only have a median overall transcription and posttranscription process8.
    [Show full text]
  • MAGE-TRIM28 Complex Promotes the Warburg Effect and Hepatocellular Carcinoma Progression by Targeting FBP1 for Degradation
    OPEN Citation: Oncogenesis (2017) 6, e312; doi:10.1038/oncsis.2017.21 www.nature.com/oncsis ORIGINAL ARTICLE MAGE-TRIM28 complex promotes the Warburg effect and hepatocellular carcinoma progression by targeting FBP1 for degradation X Jin1,2,YPan2, L Wang3, L Zhang4, R Ravichandran5, PR Potts5, J Jiang6,HWu7 and H Huang2,8,9 Hepatocellular carcinoma (HCC) is one of the leading cause of cancer death in the world. Fructose-1,6-biphosphatase (FBP1), a rate-limiting enzyme in gluconeogenesis, has been identified recently as a tumor suppressor in HCC and other cancer types. In this study, we demonstrated that the tripartite motif-containing protein 28 (TRIM28) binds directly to and promotes FBP1 for ubiquitination and degradation. MAGE-A3 and MAGE-C2, which are known to be overexpressed in HCC, can enhance TRIM28-dependent degradation of FBP1 by forming ubiquitin ligase complexes with TRIM28. We further showed that expression of TRIM28 increased glucose consumption and lactate production by promoting FBP1 degradation in HCC cells and that FBP1 is a key mediator of TRIM28-induced HCC growth in culture and in mice. Moreover, we demonstrated that FBP1 and TRIM28 protein levels inversely correlated in HCC patient specimens. Finally, we showed that the proteasome inhibitor bortezomib mitigated the Warburg effect by inhibiting FBP1 degradation in HCC. Collectively, our findings not only identify oncogenic MAGE-TRIM28 complex-mediated proteasome degradation of FBP1 as a key mechanism underlying downregulation of FBP1 proteins in HCC, but also reveal that MAGE-TRIM28-regulated reprogramming of cancer cell metabolism and HCC tumorigenesis is mediated, at least in part, through FBP1 degradation.
    [Show full text]
  • Glucose Starvation to Rapid Death of Nrf1, but Not Nrf2, Deficient Hepatoma Cells from Its Fatal Defects in the Redox Metabolism Reprogramming
    bioRxiv preprint doi: https://doi.org/10.1101/2019.12.13.875369; this version posted December 13, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Glucose starvation to rapid death of Nrf1, but not Nrf2, deficient hepatoma cells from its fatal defects in the redox metabolism reprogramming Yu-ping Zhu1, Ze Zheng1, Yuancai Xiang2, and Yiguo Zhang1* 1The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Medical Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China. 2Department of Biochemistry and Molecular Biology, Collage of Basic Medical Sciences, Southwest Medical University, Sichuan, 646000, China. *Correspondence should be addressed to Yiguo Zhang (Email: [email protected], or [email protected]) Abstract Metabolic reprogramming exists in a variety of cancer cells, with the most relevance to glucose as a source of energy and carbon for survival and proliferation. Of note, Nrf1 was shown to be essential for regulating glycolysis pathway, but it is unknown whether it plays a role in cancer metabolic reprogramming, particularly in response to glucose starvation. Herein, we discover that Nrf1α/-derived hepatoma cells are sensitive to rapid death induced by glucose deprivation, such cell death appears to be rescued by Nrf2 interference, but Nrf1/2/ or Nrf2/-derived cells are roughly unaffected by glucose starvation. Further evidence revealed that Nrf1α/ cell death is resulted from severe oxidative stress arising from abberrant redox metabolism. Strikingly, altered gluconeogenesis pathway was aggravated by glucose starvation of Nrf1α/ cells, as also accompanied by weakened pentose phosphate pathway, dysfunction of serine-to-glutathione synthesis, and accumulation of reactive oxygen species (ROS) and damages, such that the intracellular GSH and NADPH were exhausted.
    [Show full text]
  • Table S1. 103 Ferroptosis-Related Genes Retrieved from the Genecards
    Table S1. 103 ferroptosis-related genes retrieved from the GeneCards. Gene Symbol Description Category GPX4 Glutathione Peroxidase 4 Protein Coding AIFM2 Apoptosis Inducing Factor Mitochondria Associated 2 Protein Coding TP53 Tumor Protein P53 Protein Coding ACSL4 Acyl-CoA Synthetase Long Chain Family Member 4 Protein Coding SLC7A11 Solute Carrier Family 7 Member 11 Protein Coding VDAC2 Voltage Dependent Anion Channel 2 Protein Coding VDAC3 Voltage Dependent Anion Channel 3 Protein Coding ATG5 Autophagy Related 5 Protein Coding ATG7 Autophagy Related 7 Protein Coding NCOA4 Nuclear Receptor Coactivator 4 Protein Coding HMOX1 Heme Oxygenase 1 Protein Coding SLC3A2 Solute Carrier Family 3 Member 2 Protein Coding ALOX15 Arachidonate 15-Lipoxygenase Protein Coding BECN1 Beclin 1 Protein Coding PRKAA1 Protein Kinase AMP-Activated Catalytic Subunit Alpha 1 Protein Coding SAT1 Spermidine/Spermine N1-Acetyltransferase 1 Protein Coding NF2 Neurofibromin 2 Protein Coding YAP1 Yes1 Associated Transcriptional Regulator Protein Coding FTH1 Ferritin Heavy Chain 1 Protein Coding TF Transferrin Protein Coding TFRC Transferrin Receptor Protein Coding FTL Ferritin Light Chain Protein Coding CYBB Cytochrome B-245 Beta Chain Protein Coding GSS Glutathione Synthetase Protein Coding CP Ceruloplasmin Protein Coding PRNP Prion Protein Protein Coding SLC11A2 Solute Carrier Family 11 Member 2 Protein Coding SLC40A1 Solute Carrier Family 40 Member 1 Protein Coding STEAP3 STEAP3 Metalloreductase Protein Coding ACSL1 Acyl-CoA Synthetase Long Chain Family Member 1 Protein
    [Show full text]