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Expression of a Gene Encoding Acetolactate Synthase from Rice Complements Two Ilvh Mutants in Escherichia Coli
AJCS 4(6):430-436 (2010) ISSN:1835-2707 Expression of a gene encoding acetolactate synthase from rice complements two ilvH mutants in Escherichia coli Md. Shafiqul Islam Sikdar1, Jung-Sup Kim2* 1Faculty of Biotechnology, Jeju National University, Jeju, 690-756, Korea and Department of Agronomy, Hajee Mohammad Danesh Science and Technology University, Dinajpur-5200, Bangladesh 2Faculty of Biotechnology, Jeju National University, Jeju, 690-756, Korea *Corresponding Author: [email protected] Abstract Acetolactate synthase (ALS) is a thiamine diphosphate-dependent enzyme in the biosynthetic pathway leading to isoleucine, valine and leucine in plants. ALS is the target of several classes of herbicides that are effective to protect a broad range of crops. In this study, we describe the functional analysis of a gene encoding for ALS from rice (OsALS). Sequence analysis of an EST from rice revealed that it harbors a full-length open reading frame for OsALS encoding a protein of approximately 69.4 kDa and the N-terminal of OsALS contains a feature of chloroplast transit peptide. The predicted amino acid sequence of OsALS is highly homologous to those of weed ALSs among plant ALSs. The OsALS expression showed that the gene was functionally capable of complementing the two ilvH mutant strains of Escherichia coli. These results indicate that the OsALS encodes for an enzyme in acetolactate synthase in rice. Keywords: acetolactate synthase, rice (Oryza sativa), sequence analysis, functional complementation, ilvH mutants Abbreviations: ALS_Acetolactate synthase; BCAAs_Branched chain amino acids; Ile_Isoleucine; Val_Valine; Leu_Leucine; TPP_Thiamine diphosphate; CGSC_E. coli Genetic Stock Center; RGRC _Rice Genome Resource Center; ORF_Open reading frame; PCR_Polymerase chain reaction; Amp_Ampicillin; MM_M9 minimal medium; IPTG_Isopropyl β-D-thiogalactopyranoside. -
Yeast Genome Gazetteer P35-65
gazetteer Metabolism 35 tRNA modification mitochondrial transport amino-acid metabolism other tRNA-transcription activities vesicular transport (Golgi network, etc.) nitrogen and sulphur metabolism mRNA synthesis peroxisomal transport nucleotide metabolism mRNA processing (splicing) vacuolar transport phosphate metabolism mRNA processing (5’-end, 3’-end processing extracellular transport carbohydrate metabolism and mRNA degradation) cellular import lipid, fatty-acid and sterol metabolism other mRNA-transcription activities other intracellular-transport activities biosynthesis of vitamins, cofactors and RNA transport prosthetic groups other transcription activities Cellular organization and biogenesis 54 ionic homeostasis organization and biogenesis of cell wall and Protein synthesis 48 plasma membrane Energy 40 ribosomal proteins organization and biogenesis of glycolysis translation (initiation,elongation and cytoskeleton gluconeogenesis termination) organization and biogenesis of endoplasmic pentose-phosphate pathway translational control reticulum and Golgi tricarboxylic-acid pathway tRNA synthetases organization and biogenesis of chromosome respiration other protein-synthesis activities structure fermentation mitochondrial organization and biogenesis metabolism of energy reserves (glycogen Protein destination 49 peroxisomal organization and biogenesis and trehalose) protein folding and stabilization endosomal organization and biogenesis other energy-generation activities protein targeting, sorting and translocation vacuolar and lysosomal -
B Number Gene Name Mrna Intensity Mrna
sample) total list predicted B number Gene name assignment mRNA present mRNA intensity Gene description Protein detected - Membrane protein membrane sample detected (total list) Proteins detected - Functional category # of tryptic peptides # of tryptic peptides # of tryptic peptides detected (membrane b0002 thrA 13624 P 39 P 18 P(m) 2 aspartokinase I, homoserine dehydrogenase I Metabolism of small molecules b0003 thrB 6781 P 9 P 3 0 homoserine kinase Metabolism of small molecules b0004 thrC 15039 P 18 P 10 0 threonine synthase Metabolism of small molecules b0008 talB 20561 P 20 P 13 0 transaldolase B Metabolism of small molecules chaperone Hsp70; DNA biosynthesis; autoregulated heat shock b0014 dnaK 13283 P 32 P 23 0 proteins Cell processes b0015 dnaJ 4492 P 13 P 4 P(m) 1 chaperone with DnaK; heat shock protein Cell processes b0029 lytB 1331 P 16 P 2 0 control of stringent response; involved in penicillin tolerance Global functions b0032 carA 9312 P 14 P 8 0 carbamoyl-phosphate synthetase, glutamine (small) subunit Metabolism of small molecules b0033 carB 7656 P 48 P 17 0 carbamoyl-phosphate synthase large subunit Metabolism of small molecules b0048 folA 1588 P 7 P 1 0 dihydrofolate reductase type I; trimethoprim resistance Metabolism of small molecules peptidyl-prolyl cis-trans isomerase (PPIase), involved in maturation of b0053 surA 3825 P 19 P 4 P(m) 1 GenProt outer membrane proteins (1st module) Cell processes b0054 imp 2737 P 42 P 5 P(m) 5 GenProt organic solvent tolerance Cell processes b0071 leuD 4770 P 10 P 9 0 isopropylmalate -
The Influence of Cell Cycle Regulation on Chemotherapy
International Journal of Molecular Sciences Review The Influence of Cell Cycle Regulation on Chemotherapy Ying Sun 1, Yang Liu 1, Xiaoli Ma 2 and Hao Hu 1,* 1 Institute of Biomedical Materials and Engineering, College of Materials Science and Engineering, Qingdao University, Qingdao 266071, China; [email protected] (Y.S.); [email protected] (Y.L.) 2 Qingdao Institute of Measurement Technology, Qingdao 266000, China; [email protected] * Correspondence: [email protected] Abstract: Cell cycle regulation is orchestrated by a complex network of interactions between proteins, enzymes, cytokines, and cell cycle signaling pathways, and is vital for cell proliferation, growth, and repair. The occurrence, development, and metastasis of tumors are closely related to the cell cycle. Cell cycle regulation can be synergistic with chemotherapy in two aspects: inhibition or promotion. The sensitivity of tumor cells to chemotherapeutic drugs can be improved with the cooperation of cell cycle regulation strategies. This review presented the mechanism of the commonly used chemotherapeutic drugs and the effect of the cell cycle on tumorigenesis and development, and the interaction between chemotherapy and cell cycle regulation in cancer treatment was briefly introduced. The current collaborative strategies of chemotherapy and cell cycle regulation are discussed in detail. Finally, we outline the challenges and perspectives about the improvement of combination strategies for cancer therapy. Keywords: chemotherapy; cell cycle regulation; drug delivery systems; combination chemotherapy; cancer therapy Citation: Sun, Y.; Liu, Y.; Ma, X.; Hu, H. The Influence of Cell Cycle Regulation on Chemotherapy. Int. J. 1. Introduction Mol. Sci. 2021, 22, 6923. https:// Chemotherapy is currently one of the main methods of tumor treatment [1]. -
The Effect of Increased Plastid Transketolase Activity on Thiamine
The effect of increased plastid transketolase activity on thiamine metabolism in transgenic tobacco plants Stuart James Fisk A thesis submitted for the degree of Doctor of Philosophy School of Biological Sciences University of Essex November 2015 I Acknowledgements I'd like to thank my supervisors Professor Christine Raines and Dr Tracy Lawson for all of the support and encouragement, Dr Julie Lloyd for the comments and suggestions given at board meetings and the University of Essex for funding this research. An extremely big thank you to all the members of plant group both past and present who have provided lots of help and technical support as well as plenty of laughs and an addiction to caffeine. But the biggest thanks of all go to my wife and best friend Alison Fisk. She always believed in me. II Summary Transketolase is a TPP dependent enzyme that affects the availability of intermediates in both the Calvin cycle and non-oxidative pentose phosphate pathway. Previous studies have indicated that changes to the activity level of transketolase can limit growth and development as well as the production of isoprenoids, starch, amino acids and thiamine. The overall aim of this project was to further advance the understanding of the mechanism linking increased TK activity and thiamine metabolism. Nicotiana tabacum mutants with increased total transketolase activity ~ 2 to 2.5 fold higher than WT plants were shown to have a reduced growth and chlorotic phenotype. In seedlings, these phenotypes were attributed to a reduction in seed thiamine content. Imbibition of TKox seeds in a thiamine solution produced plants that were comparable to WT plants. -
The Role of Sumoylation of DNA Topoisomerase Iiα C-Terminal Domain in the Regulation of Mitotic Kinases In
SUMOylation at the centromere: The role of SUMOylation of DNA topoisomerase IIα C-terminal domain in the regulation of mitotic kinases in cell cycle progression. By Makoto Michael Yoshida Submitted to the graduate degree program in the Department of Molecular Biosciences and the Graduate Faculty of the University of Kansas in partial fulfillment of the requirements for the degree of Doctor of Philosophy. ________________________________________ Chairperson: Yoshiaki Azuma, Ph.D. ________________________________________ Roberto De Guzman, Ph.D. ________________________________________ Kristi Neufeld, Ph.D. _________________________________________ Berl Oakley, Ph.D. _________________________________________ Blake Peterson, Ph.D. Date Defended: July 12, 2016 The Dissertation Committee for Makoto Michael Yoshida certifies that this is the approved version of the following dissertation: SUMOylation at the centromere: The role of SUMOylation of DNA topoisomerase IIα C-terminal domain in the regulation of mitotic kinases in cell cycle progression. ________________________________________ Chairperson: Yoshiaki Azuma, Ph.D. Date approved: July 12, 2016 ii ABSTRACT In many model systems, SUMOylation is required for proper mitosis; in particular, chromosome segregation during anaphase. It was previously shown that interruption of SUMOylation through the addition of the dominant negative E2 SUMO conjugating enzyme Ubc9 in mitosis causes abnormal chromosome segregation in Xenopus laevis egg extract (XEE) cell-free assays, and DNA topoisomerase IIα (TOP2A) was identified as a substrate for SUMOylation at the mitotic centromeres. TOP2A is SUMOylated at K660 and multiple sites in the C-terminal domain (CTD). We sought to understand the role of TOP2A SUMOylation at the mitotic centromeres by identifying specific binding proteins for SUMOylated TOP2A CTD. Through affinity isolation, we have identified Haspin, a histone H3 threonine 3 (H3T3) kinase, as a SUMOylated TOP2A CTD binding protein. -
Mining the Arabidopsis Genome for Cytochrome P450 Biocatalysts
Mining the Arabidopsis genome for cytochrome P450 biocatalysts Maria Magdalena Razalan PhD University of York Biology September 2016 Abstract Cytochromes P450 (CYPs) constitute a wide group of NAD(P)H-dependent monooxygenases, found throughout all kingdoms of life. Among the most important functions of CYPs are the synthesis of bioactive compounds and the conversion of xenobiotics. These functions can be translated into biotechnological applications, such as the production of highly regio- and stereo-specific drug metabolites for the pharmaceutical industry, or to confer activity towards toxic compounds for agronomics and bioremediation purposes. Plants possess a large number of CYP sequences, but most still remain uncharacterised, due to the difficulty in the isolation of the membrane-bound enzyme and in the reconstitution of an active and efficient redox system. In this project, a fusion construct for the co-expression of the CYP with a suitable reductase was created. The construct consisted of a C-terminal Arabidopsis ATR2 reductase (codon-optimised for expression in E. coli and truncated of the N-terminal membrane anchor) connected through a poly-GlySer linker to the heme domain. An N-terminal Im9 peptide replaced the natural membrane-binding domain of the CYP. When CYP73A5 from Arabidopsis was cloned into the construct, it was able to convert almost 60 % of the substrate cinnamic acid to the hydroxylated derivative, in whole cell assays. This result demonstrated that this expression platform enables the expression of active redox self-sufficient P450 catalysts and it can be further utilised for the characterisation of orphan CYPs. Following from gene expression studies and reports on the existence of oxidative derivatives of TNT, the potential involvement of CYP81D11 in the detoxification of TNT was explored with different in planta assays, employing transgenic Arabidopsis lines and tobacco leaf discs. -
Ciprofloxacin Stress Proteome of the Extended-Spectrum Beta-Lactamase
Ciprofloxacin stress proteome of the extended-spectrum beta-lactamase producing Escherichia coli from slaughtered pigs Sónia Ramos, Ingrid Chafsey, Michel Hébraud, Margarida Sousa, Patrícia Poeta, Gilberto Igrejas To cite this version: Sónia Ramos, Ingrid Chafsey, Michel Hébraud, Margarida Sousa, Patrícia Poeta, et al.. Ciprofloxacin stress proteome of the extended-spectrum beta-lactamase producing Escherichia coli from slaughtered pigs. Current Proteomics, Bentham Science Publishers, 2016, 13, pp.285-289. 10.2174/1570164613666161018144215. hal-01602438 HAL Id: hal-01602438 https://hal.archives-ouvertes.fr/hal-01602438 Submitted on 27 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Send Orders for Reprints to [email protected] Current Proteomics, 2016, 13, 1-5 1 RESEARCH ARTICLE Ciprofloxacin Stress Proteome of the Extended-Spectrum -lactamase Producing Escherichia coli from Slaughtered Pigs Sónia Ramos1,2,3,4, Ingrid Chafsey5, Michel Hébraud5,6, Margarida Sousa1,2,3,4, Patrícia Poeta4,7 and Gilberto Igrejas1,2,7,* 1Functional Genomics -
In-Silico Three Dimensional Structure Prediction of Important Neisseria Meningitidis Proteins
doi.org/10.36721/PJPS.2021.34.2.REG.553-560.1 In-silico three dimensional structure prediction of important Neisseria meningitidis proteins Muhammad Ali1, Muhammad Aurongzeb2 and Yasmeen Rashid1* 1Department of Biochemistry, University of Karachi, Karachi, Pakistan 2Jamil-ur-Rehman Center for Genome Research, PCMD, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan Abstract: Pathogenic bacteria Neisseria meningitidis cause serious infection i.e. meningitis (infection of the brain) worldwide. Among five pathogenic serogroups, serogroup B causes life threatening illness as there is no effective vaccine available due to its poor immunogenicity. A total of 73 genes in N. meningitidis genome have identified that were proved to be essential for meningococcal disease and were considered as crucial drug targets. We targeted five of those proteins, which are known to involve in amino acid biosynthesis, for homology-based three dimensional structure determinations by MODELLER (v9.19) and evaluated the models by PROSA and PROCHECK programs. Detailed structural analyses of NMB0358, NMB0943, NMB1446, NMB1577 and NMB1814 proteins were carried out during the present research. Based on a high degree of sequence conservation between target and template protein sequences, excellent models were built. The overall three dimensional architectures as well as topologies of all the proteins were quite similar with that of the templates. Active site residues of all the homology models were quite conserved with respect to their respective templates indicating similar catalytic mechanisms in these orthologues. Here, we are reporting, for the first time, detailed three dimensional folds of N. meningitidis pathogenic factors involved in a crucial cellular metabolic pathway. -
Characterization of Acetolactate Synthase Resistance in Common Sunflower (Helianthus Annuus) Anthony D
Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2002 Characterization of acetolactate synthase resistance in common sunflower (Helianthus annuus) Anthony D. White Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Agricultural Science Commons, Agriculture Commons, and the Agronomy and Crop Sciences Commons Recommended Citation White, Anthony D., "Characterization of acetolactate synthase resistance in common sunflower (Helianthus annuus) " (2002). Retrospective Theses and Dissertations. 407. https://lib.dr.iastate.edu/rtd/407 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g.. maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. -
A Protein Kinase Activity Tightly Associated with Drosophila Type II
Proc. Nati. Acad. Sci. USA Vol. 81, pp. 6938-6942, November 1984 Biochemistry A protein kinase activity tightly associated with Drosophila type II DNA topoisomerase (eukaryotic topoisomerase/protein phosphorylation/Drosophila melanogaster) MIRIAM SANDER, JAMES M. NOLAN, AND TAO-SHIH HSIEH Department of Biochemistry, Duke University Medical Center, Durham, NC 27710 Communicated by Robert L. Hill, July 19, 1984 ABSTRACT A protein kinase activity has been identified tial gene. The mechanism by which the eukaryotic topoiso- that is tightly associated with the purified Drosophila type II merase activities are regulated is not yet clear. Since varia- DNA topoisomerase. The kinase and topoisomerase activities tions in chromatin structure are associated with the are not separated when the enzyme is subjected to analytical processes of transcription, replication, and cell division, it is chromatography (phosphoceilulose, single-strand DNA aga- likely that topoisomerase-mediated alteration in chromatin rose, and Sephacryl S-300) and analytical glycerol gradient structure will be regulated in a complex manner. sedimentation. These two activities are also inactivated to the Recently, a type I topoisomerase isolated from Novikoff same extent by either heat or N-ethylmaleimide treatment. hepatoma cells has been shown to be a phosphoprotein (15). The evidence, however, does not rule out the possibility that In our studies on Drosophila type II topoisomerase we have the kinase activity resides in a polypeptide other than the to- established that the enzyme isolated from the Kco Drosophi- poisomerase polypeptide. The topoisomerase-associated pro- la cell culture line is present in a phosphorylated form (un- tein kinase activity is not stimulated by Ca21 or cyclic nucleo- published data). -
Direct Control of Type IIA Topoisomerase Activity by a Chromosomally Encoded Regulatory Protein
Direct control of type IIA topoisomerase activity by a chromosomally encoded regulatory protein Seychelle M. Vos,1,4 Artem Y. Lyubimov,1,5 David M. Hershey,2 Allyn J. Schoeffler,1,6 Sugopa Sengupta,3,7 Valakunja Nagaraja,3 and James M. Berger1,8,9 1Department of Molecular and Cell Biology, 2Deparment of Plant and Microbial Biology, University of California at Berkeley, Berkeley, California 94720, USA; 3Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560 012, India Precise control of supercoiling homeostasis is critical to DNA-dependent processes such as gene expression, replication, and damage response. Topoisomerases are central regulators of DNA supercoiling commonly thought to act independently in the recognition and modulation of chromosome superstructure; however, recent evidence has indicated that cells tightly regulate topoisomerase activity to support chromosome dynamics, transcriptional response, and replicative events. How topoisomerase control is executed and linked to the internal status of a cell is poorly understood. To investigate these connections, we determined the structure of Escherichia coli gyrase, a type IIA topoisomerase bound to YacG, a recently identified chromosomally encoded inhibitor protein. Phylogenetic analyses indicate that YacG is frequently associated with coenzyme A (CoA) production enzymes, linking the protein to metabolism and stress. The structure, along with supporting solution studies, shows that YacG represses gyrase by sterically occluding the principal DNA-binding site of the enzyme. Unexpectedly, YacG acts by both engaging two spatially segregated regions associated with small-molecule inhibitor interactions (fluoroquinolone antibiotics and the newly reported antagonist GSK299423) and remodeling the gyrase holoenzyme into an inactive, ATP-trapped configuration.