Digital Commons Network^™

Hyaluronate Lyase Gene Of Streptococcus Pyogenes: Molecular Characterization And Regulation, Sheryl Lynne Walton

Theses and Dissertations in Biomedical Sciences

Streptococcus pyogenes causes an assortment of diseases ranging from pharyngitis to necrotizing fasciitis. This bacterium has the ability to elaborate several extracellular products capable of causing tissue damage; one of which is a hyaluronate lyase. Little information is available regarding the regulation of streptococcal hyaluronidase. As such, the expression of hylA in the hyaluronate lyase-producing strain ATCC 10403 and in strains that do not produce detectable enzyme was analyzed by RT-PCR and quantitative PCR. Hyaluronate lyase transcript was detected throughout growth for all strains; however, expression of hylA in the enzymatically inactive strains, 71698 and SF370, was determined …

The Quantitative Assessment Of Osteoinductivity Of Human Demineralized Bone Matrix And Cdna Array Analysis Of Osteogenic Differentiation In Human Periosteal Cells, Sittisak Honsawek

Theses and Dissertations in Biomedical Sciences

Demineralized bone matrix (DBM) has been extensively utilized in orthopeadic, periodontal, and maxillofacial applications. DBM comprises bone morphogenetic proteins (BMPs) that are essential regulators for endochondral bone formation. The current study was undertaken to establish the quantitative in vitro assays for determining the osteoinductive potential of human DBM and the use of cDNA gene expression array technology for examining gene expression profiles during osteoblast differentiation in human periosteal cells.

BMP-4 is one of the most osteoinductive factors and has been investigated for clinical applications. For this reason, BMP-4 was designated as the osteoinductive protein for use in the quantitative in …

Emissions Of Trace Gases And Particles From Savanna Fires In Southern Africa, Parikhit Sinha, Peter V. Hobbs, Robert J. Yokelson, I. T. Bertschi, Donald R. Blake, Isobel J. Simpson, Song Gao, Thomas W. Kirschsetter, Tica Novakov

Chemistry and Biochemistry Faculty Publications

[1] Airborne measurements made on initial smoke from 10 savanna fires in southern Africa provide quantitative data on emissions of 50 gaseous and particulate species, including carbon dioxide, carbon monoxide, sulfur dioxide, nitrogen oxides, methane, ammonia, dimethyl sulfide, nonmethane organic compounds, halocarbons, gaseous organic acids, aerosol ionic components, carbonaceous aerosols, and condensation nuclei (CN). Measurements of several of the gaseous species by gas chromatography and Fourier transform infrared spectroscopy are compared. Emission ratios and emission factors are given for eight species that have not been reported previously for biomass burning of savanna in southern Africa (namely, dimethyl sulfide, methyl nitrate, …

Role Of Proapoptotic Bax In Propagation Of Chlamydia Muridarum (The Mouse Pneumonitis Strain Of Chlamydia Trachomatis) And The Host Inflammatory Response, Jean-Luc Perfettini, David M. Ojcius, Charles W. Andrews Jr., Stanley J. Korsmeyer, Roger G. Rank, Toni Darville

All Dugoni School of Dentistry Faculty Articles

The BCL-2 family member BAX plays a critical role in regulating apoptosis. Surprisingly, bax-deficient mice display limited phenotypic abnormalities. Here we investigate the effect of BAX on infection by the sexually transmitted pathogen,Chlamydia muridarum (the mouse pneumonitis strain ofChlamydia trachomatis). Bax −/−cells are relatively resistant to Chlamydia-induced apoptosis, and fewer bacteria are recovered after two infection cycles from Bax −/− cells than from wild-type cells. These results suggest that BAX-dependent apoptosis may be used to initiate a new round of infection, most likely by releasingChlamydia-containing apoptotic bodies from infected cells that could be internalized by neighboring uninfected cells. Nonetheless, infected …

Purified Particulate Methane Monooxygenase From Methylococcus Capsulatus (Bath) Is A Dimer With Both Mononuclear Copper And A Copper-Containing Cluster, Raquel L. Lieberman, Deepak B. Shrestha, Peter E. Doan, Brian M. Hoffman, Timothy L. Stemmler, Amy C. Rosenzweig

Biochemistry and Molecular Biology Faculty Publications

Particulate methane monooxygenase (pMMO) is a membrane-bound enzyme that catalyzes the oxidation of methane to methanol in methanotropic bacteria. Understanding how this enzyme hydroxylates methane at ambient temperature and pressure is of fundamental chemical and potential commercial importance. Difficulties in solubilizing and purifying active pMMO have led to conflicting reports regarding its biochemical and biophysical properties, however. We have purified pMMO from Methylococcus capsulatus (Bath) and detected activity. The purified enzyme has a molecular mass of ~200 kDa, probably corresponding to an a2b2g2 polypeptide arrangement. Each 200 kDa pMMO complex contains 4.8 ± 0.8 copper ions and 1.5 ± 0.7 …

Characterization Of A Gene Encoding Two Isoforms Of A Mitochondrial Protein Upregulated By Cyclosporin A In Activated T Cells, Laurent Mascarell, Rodolphe Auger, Andres Alcover, David M. Ojcius, Thomas Jungas, Veronique Cadet-Daniel, Jean M. Kanellopoulos, Paolo Truffa-Bacchi

All Dugoni School of Dentistry Faculty Articles

Cyclosporin A (CSA) is an immunosuppressor used in organ transplantation. A recent proteomic analysis has revealed that activation of T cells in the presence of CSA induces the synthesis of hundreds of new proteins. Here we used representational difference analysis to characterize some of the corresponding induced genes. After cDNA bank screening we focused on one of these genes, which we named CSA-conditional, T cell activation-dependent (CSTAD) gene. This gene produces two mRNAs resulting from alternative splicing events. They encode two proteins of 104 and 141 amino acids, CSTADp-S and CSTADp-L, for the short and long forms, respectively. FK506 had …

Evolution Of Gases And Particles From A Savanna Fire In South Africa, Peter V. Hobbs, Parikhit Sinha, Robert J. Yokelson, Ted J. Christian, Donald R. Blake, Song Gao, Thomas W. Kirschsetter, Tica Novakov, Peter Pilewskie

Chemistry and Biochemistry Faculty Publications

[1] Airborne measurements of particles and gases from a 1000-ha savanna fire in South Africa are presented. These measurements represent the most extensive data set reported on the aging of biomass smoke. The measurements include total concentrations of particles (CN), particle sizes, particulate organic carbon and black carbon, light-scattering coefficients, downwelling UV fluxes, and mixing ratios for 42 trace gases and 7 particulate species. The ratios of excess nitrate, ozone, and gaseous acetic acid to excess CO increased significantly as the smoke aged over ∼40–45 min, indicating that these species were formed by photochemistry in the plume. For 17 other …

Androgen-Induced Recruitment Of Rna Polymerase Ii To A Nuclear Receptor–P160 Coactivator Complex, Maggie C. Louie, Hong Qiong Yang, Ai-Hong Ma, Wei Xu, June X. Zou, Hsing-Jien Kung, Hong-Wu Chen

Collected Faculty Scholarship

The androgen receptor, like other nuclear receptors, activates target genes by binding to hormone-responsive enhancers. Here we demonstrate that androgen induces robust recruitment of androgen receptor, members of the p160 coactivator family, and CREB-binding protein/p300 specifically at the distant enhancer of prostate-specific antigen (PSA) gene. Unexpectedly, we found that RNA polymerase II (Pol II) is directly recruited to the enhancer in a hormone-dependent manner, independent of the proximal promoter, and that the isolated PSA enhancer can mediate efficient androgen induction of transcription. Inhibition of the Pol II carboxyl-terminal domain kinase activity with low concentrations of flavopiridol blocks Pol II transfer …

Minus-End Capture Of Preformed Kinetochore Fibers Contributes To Spindle Morphogenesis, Alexey Khodjakov, Lily Copenagle, Michael B. Gordon, Duane A. Compton, Tarun M. Kapoor

Dartmouth Scholarship

Near-simultaneous three-dimensional fluorescence/differential interference contrast microscopy was used to follow the behavior of microtubules and chromosomes in living alpha-tubulin/GFP-expressing cells after inhibition of the mitotic kinesin Eg5 with monastrol. Kinetochore fibers (K-fibers) were frequently observed forming in association with chromosomes both during monastrol treatment and after monastrol removal. Surprisingly, these K-fibers were oriented away from, and not directly connected to, centrosomes and incorporated into the spindle by the sliding of their distal ends toward centrosomes via a NuMA-dependent mechanism. Similar preformed K-fibers were also observed during spindle formation in untreated cells. In addition, upon monastrol removal, centrosomes established a transient …

Selective Removal Of The Selenocysteine Trna [Ser]Sec Gene (Trsp) In Mouse Mammary Epithelium, Easwari Kumaraswamy, Bradley A. Carlson, Fanta Morgan, Keiko Miyoshi, Gertraud W. Robinson, Dan Su, Shulin Wang, Eileen Southon, Lino Tessarollo, Byeong Jae Lee, Vadim Gladyshev, Lothar Hennighausen, Dolph L. Hatfield

Vadim Gladyshev Publications

Mice homozygous for an allele encoding the selenocysteine (Sec) tRNA ^[Ser]Sec gene (Trsp) flanked by loxP sites were generated. Cre recombinase-dependent removal of Trsp in these mice was lethal to embryos. To investigate the role of Trsp in mouse mammary epithelium, we deleted this gene by using transgenic mice carrying the Cre recombinase gene under control of the mouse mammary tumor virus (MMTV) long terminal repeat or the whey acidic protein promoter. While both promoters target Cre gene expression to mammary epithelium, MMTV-Cre is also expressed in spleen and skin. Sec tRNA ^[Ser]Sec amounts were reduced by …

Enzymatic Method For Assaying Calcium In Serum With Ca++ -Atpase, Masood-Ul-Hassan Javed, Francesco Michelangeli

Department of Biological & Biomedical Sciences

A kinetic assay for total calcium in serum was developed which is based on the activation of Ca(++)-ATPase by free Ca(++) [Ca(++)](f) maintained by EGTA in the reaction mixture. The concentration of Ca(++)(f) was dependent on total reference calcium added or serum calcium. Ca(++)-ATPase activity was coupled to the reduction of NADH by pyruvate kinase (PK) and lactate dehydrogenase (LDH) and monitored by change in absorbance at 340 nm. The calcium in normal serum was 10.08 +/- 0.24 mg/dl (n = 35) by our method while with o-cresolphthalein complexone (CPC) method, the total calcium in the same 35 serum samples …

Rpb4p, A Subunit Of Rna Polymerase Ii, Mediates Mrna Export During Stress, Marganit Farago, Tal Nahari, Christopher Hammel, Charles N. Cole, Mordechai Choder

Dartmouth Scholarship

Changes in gene expression represent a major mechanism by which cells respond to stress. We and other investigators have previously shown that the yeast RNA polymerase II subunit Rpb4p is required for transcription under various stress conditions, but not under optimal growth conditions. Here we show that, in addition to its role in transcription, Rpb4p is also required for mRNA export, but only when cells are exposed to stress conditions. The roles of Rpb4p in transcription and in mRNA export can be uncoupled genetically by specific mutations in Rpb4p. Both functions of Rpb4p are required to maintain cell viability during …

Trace Gas And Particle Emissions From Fires In Large Diameter And Belowground Biomass Fuels, Isaac T. Bertschi, Robert J. Yokelson, Darold E. Ward, R. E. Babbitt, Ronald A. Susott, Jon G. Goode, Wei Min Hao

Chemistry and Biochemistry Faculty Publications

[1] We adopt a working definition of residual smoldering combustion (RSC) as biomass combustion that produces emissions that are not lofted by strong fire-induced convection. RSC emissions can be produced for up to several weeks after the passage of a flame front and they are mostly unaffected by flames. Fuels prone to RSC include downed logs, duff, and organic soils. Limited observations in the tropics and the boreal forest suggest that RSC is a globally significant source of emissions to the troposphere. This source was previously uncharacterized. We measured the first emission factors (EF) for RSC in a series of …

Examination Of The Role Of The Clamp-Loader And Atp Hydrolysis In The Formation Of The Bacteriophage T4 Polymerase Holoenzyme, Michael A. Trakselis, Anthony J. Berdis, Stephen J. Benkovic

Chemistry Faculty Publications

Transient kinetic analyses further support the role of the clamp-loader in bacteriophage T4 as a catalyst which loads the clamp onto DNA through the sequential hydrolysis of two molecules of ATP before and after addition of DNA. Additional rapid-quench and pulse-chase experiments have documented this stoichiometry. The events of ATP hydrolysis have been related to the opening/closing of the clamp protein through fluorescence resonance energy transfer (FRET). In the absence of a hydrolysable form of ATP, the distance across the subunit interface of the clamp does not increase as measured by intramolecular FRET, suggesting gp45 cannot be loaded onto DNA. …

Trace Gas Emissions From The Production And Use Of Domestic Biofuels In Zambia Measured By Open-Path Fourier Transform Infrared Spectroscopy, Isaac T. Bertschi, Robert J. Yokelson, Darold E. Ward, Ted J. Christian, Wei Min Hao

Chemistry and Biochemistry Faculty Publications

[1] Domestic biomass fuels (biofuels) were recently estimated to be the second largest source of carbon emissions from global biomass burning. Wood and charcoal provide approximately 90% and 10% of domestic energy in tropical Africa. In September 2000, we used open-path Fourier transform infrared (OP-FTIR) spectroscopy to quantify 18 of the most abundant trace gases emitted by wood and charcoal cooking fires and an earthen charcoal-making kiln in Zambia. These are the first in situ measurements of an extensive suite of trace gases emitted by tropical biofuel burning. We report emission ratios (ER) and emission factors (EF) for (in order …

Phosphoproteomic Studies Of Smooth Muscle Contraction: Investigation Of Differential Phosphorylation In Relaxed/Contracted Rat Aortic Smooth Muscle Tissue Using Maldi-Tof Ms, Tonya M. Pekar

Theses, Dissertations and Capstones

Many human disorders are associated with the malfunction of smooth muscle tissue, or are related to the capabilities of its proper function—asthma, glaucoma, renal inefficiency, hypertension, and cardiovascular disease. Dysfunctional proteins are frequently implicated as the source of such disorders. As the second highest cause of death in the United States, the epidemic of cardiovascular disease makes the study of smooth muscle of utmost concern.

The capabilities of proteomics and mass spectrometry allow the entire proteome complement of a cell or tissue type to be analyzed at once. This investigation employs such techniques in an effort to better understand the …

Effect Of Dimethyl Sulfoxide And Dimethylformamide On The Stability Of 4-Halo-8quinolinols / Herman Gershon, Donald D. Clarke, And John J. Mcmahon Department Of Chemistry, Fordham University, Bronx, Ny 10458-9993, Usa New York Botanical Garden, Bronx, Ny 10458-9993, Usa, Herman Gershon, Donald Dudley Clarke Phd, John J. Mcmahon

Chemistry Faculty Publications

Polyhalo-8-quinolinols with chlorine or bromine in position 4 were not stable in DMSO or DMF. The degradation product from 4,5-dichloro-8-quinolinol was 5-chloro-4,8-quinolindiol and the major product from 4,5-dibromo-8-quinolinol was 3,5-dibromo-4,8-quinolindiol. 4,5,7-Trichloro- and 4,5,7-tribromo-8-quinolinols yielded similar hydrolytic products, and for the bromo compound, a rebrominated product in DMSO. In DMF rebrornination did not occur. In pyridine-d5 these reactions did not take place, indicating a special ability of DMSO and DMF to cause such hydrolysis at position 4 of 4-halo-8-quinolinols

Tetratricopeptide Repeat Motif-Mediated Hsc70-Msti1 Interaction Molecular Characterization Of The Critical Contacts For Successful Binding And Specificity*, Odutayo O. Odunuge, Judith A. Hornby, Chrisiane Bies, Richard Zimmermann, David J. Pugh, Gregory L. Blatch

Faculty Publications

Murine stress-inducible protein 1 (mSTI1) is a cochaperone that is homologous with the human Hsp70/ Hsp90-organizing protein (Hop). Guided by Hop structural data and sequence alignment analyses, we have used site-directed mutagenesis, co-precipitation assays, circular dichroism spectroscopy, steady-state fluorescence, and surface plasmon resonance spectroscopy to both qualitatively and quantitatively characterize the contacts necessary for the N-terminal tetratricopeptide repeat domain (TPR1) of mSTI1 to bind to heat shock cognate protein 70 (Hsc70) and to discriminate between Hsc70 and Hsp90. We have shown that substitutions in the first TPR motif of Lys8 or Asn12 did not affect binding of mSTI1 to Hsc70, …

Fluorescence Determination Of Tryptophan Side-Chain Accessibility And Dynamics In Triple-Helical Collagen-Like Peptides, K. V. Simon-Lukasik, A. V. Persikov, B. Brodsky, J. A. M. Ramshaw, William Laws, J. B. A. Ross, R. D. Ludescher

Chemistry and Biochemistry Faculty Publications

We report tryptophan fluorescence measurements of emission intensity, iodide quenching, and anisotropy that describe the environment and dynamics at X and Y sites in stable collagen-like peptides of sequence (Gly-X-Y)(n). About 90% of tryptophans at both sites have similar solvent exposed fluorescence properties and a lifetime of 8.5-9 ns. Analysis of anisotropy decays using an associative model indicates that these long lifetime populations undergo rapid depolarizing motion with a 0.5 ns correlation time; however, the extent of fast motion at the Y site is considerably less than the essentially unrestricted motion at the X site. About 10% of tryptophans at …

Constrained Analysis Of Fluorescence Anisotropy Decay: Application To Experimental Protein Dynamics, E. Feinstein, G. Deikus, E. Rusinova, E. L. Rachofsky, J. B. A. Ross, William Laws

Chemistry and Biochemistry Faculty Publications

Hydrodynamic properties as well as structural dynamics of proteins can be investigated by the well-established experimental method of fluorescence anisotropy decay. Successful use of this method depends on determination of the correct kinetic model, the extent of cross-correlation between parameters in the fitting function, and differences between the timescales; of the depolarizing motions and the fluorophore's fluorescence lifetime. We have tested the utility of an independently measured steady-state anisotropy value as a constraint during data analysis to reduce parameter cross correlation and to increase the timescales over which anisotropy decay parameters can be recovered accurately for two calcium-binding proteins. Mutant …

Methylene Tetrahydrofolate Reductase Gene And Coronary Artery Disease, M P. Iqbal, P M. Frossard

Department of Biological & Biomedical Sciences

Abstract is not provided by the author/publisher

Renal Cortical Necrosis: A Case Series Of Nine Patients & Review Of Literature, Syed Sohail Ali, Syed Zulfiqar Rizvi, Suhail Muzaffar, Aasim Ahmad, Akhtar Ali, Sheema H. Hassan

Department of Biological & Biomedical Sciences

Background: The purpose of this study is to review cases of renal cortical necrosis (RCN) which we came across during a period of five years, confirmed on needle biopsy specimens, with literature available so far.
Methods: All renal biopsy records between January 1996 to December 2001 at the Aga Khan University Hospital, Karachi, Pakistan were reviewed to identify patients with histologically proven renal cortical necrosis. Seven patients were also referred by the department of Nephrology, Lady Reading Hospital, Peshawar, Pakistan. Patients' medical records were reviewed for demographic, clinical, and follow up data.
Results: There were total of nine patients identified …

Rec12 (Spo11) Recominase Of Fission Yeast Promotes A Backup, Distributive Pathway For Chromosome Segregation In Meiosis I, Wallace D. Sharif, Mari K. Davidson, Wayne P. Wahls

Journal of the Arkansas Academy of Science

We studied the relationship between recombination and segregation in the fission yeast Schizosaccharomyces pombe. In meiosis, chromosomes undergo two rounds of segregation to produce haploid meiotic products. Crossover meiotic ecombination (chiasmata) promotes chromosome segregation during meiosis I; achiasmatic chromosomes often suffer on disjunction in meiosis I. Recl2 protein is a topoisomerase IIortholog that introduces double-strand DNA breaks that nitiate recombination. The red2 null (deletion) and active site (Y98F) mutants lack recombination and crossovers, and onsequently suffer meiosis I nondisjunction. However, null mutants chromosomes segregate to opposite poles more frequently than predicted. Thus, fission yeast has a backup, "distributive segregation" pathway …

Isovolumetric Regulation In Mammal Cells: Role Of Taurine, B. Ordaz, R. Franco, National University Of Mexico Tuz

School of Veterinary and Biomedical Sciences: Faculty Publications

The ability to regulate cell volume is an ancient conserved trait present in essentially all species through evolution. The maintenance of a constant cell volume is a homeostatic imperative in animal cells. Changes in cell water content affecting the concentration of intracellular messenger molecules impair the complex signaling network, crucial for cell functioning and intercellular communication. Although the renal homeostatic mechanisms exert a precise control of extracellular fluid osmolarity, this is challenged in a variety of pathological situations. The intracellular volume constancy is continuously compromised by the generation of local and transient osmotic microgradients, associated with nutrients uptake, secretion, cytoskeleton …

Demographic Assessment Of The Blue Crab (Callinectes Sapidus) In Chesapeake Bay Using Extractable Lipofuscins As Age Markers, Se-Jong Ju, David H. Secor, H. Rodger Harvey

OES Faculty Publications

The blue crab (Callinectes sapidus) plays an important economic and ecological role in estuaries and coastal habitats from the Gulf of Mexico to the east coast of North America, but demographic assessments are limited by length-based methods. We applied an alternative aging method using biochemical measures of metabolic byproducts (lipofuscins) sequestered in the neural tissue of eyestalks to examine population age structure. From Chesapeake Bay, subsamples of animals collected from the 1998-99 (n-769) and 1999-2000 (n=367) winter dredge surveys were collected and lipofuscin was measured. Modal analysis of the lipofuscin index provided separation into three modes, whereas carapace-width …

Protocols For Disease Classification From Mass Spectrometry Data, Michael Wagner, Dayanand Naik, Alex Pothen

Mathematics & Statistics Faculty Publications

We report our results in classifying protein matrix-assisted laser desorption/ionizationtime of flight mass spectra obtained from serum samples into diseased and healthy groups. We discuss in detail five of the steps in preprocessing the mass spectral data for biomarker discovery, as well as our criterion for choosing a small set of peaks for classifying the samples. Cross-validation studies with four selected proteins yielded misclassification rates in the 10-15% range for all the classification methods. Three of these proteins or protein fragments are down-regulated and one up-regulated in lung cancer, the disease under consideration in this data set. When cross-validation studies …

Historical Perspectives (Apoptosis), James Curtin, Thomas Cotter

Articles

Apoptosis is one of the most widely studied fields in biology and accounts for over 2% of all life science publications annually. It plays a fundamental role in development, and defects in the regulation of apoptosis are directly implicated in numerous well-known diseases including cancer, neurodegenerative disorders, tissue atrophy and auto-immune diseases. However, the field of apoptosis has humble beginnings and was neglected by biologists for much of its history. This chapter reviews the history of research in apoptosis and highlights key experiments that have contributed significantly to our current understanding of apoptosis. In addition, the topics covered in later …

Urer, The Transcriptional Activator Of The Proteus Mirabilis Urease Gene Cluster, Is Required For Urease Activity And Virulence In Experimental Urinary Tract Infections, Jonathan D. Dattelbaum, C. Virginia Lockatell, David E. Johnson, Harry L.T. Mobley

Chemistry Faculty Publications

Proteus mirabilis, a cause of complicated urinary tract infection, produces urease, an essential virulence factor for this species. UreR, a member of the AraC/XylS family of transcriptional regulators, positively activates expression of the ure gene cluster in the presence of urea. To specifically evaluate the contribution of UreR to urease activity and virulence in the urinary tract, a ureR mutation was introduced into P. mirabilis HI4320 by homologous recombination. The isogenic ureR::aphA mutant, deficient in UreR production, lacked measurable urease activity. Expression was not detected in the UreR-deficient strain by Western blotting with monoclonal antibodies raised against UreD. Urease …

Osmosensitive Taurine Release: Does Taurine Share The Same Efflux Pathway With Chloride And Other Amino Acid Osmolytes?, Rodrigo Franco

School of Veterinary and Biomedical Sciences: Faculty Publications

Swelling subsequent to hyposmotic conditions activates a process of volume regulation present in most cell types. This volume adjustment is accomplished by osmolyte translocation toward the extracellular space to reach a new osmotic equilibrium. Molecules involved in this homeostatic mechanism have been broadly classified into two categories: organic and inorganic osmolytes. Inorganic osmolytes comprise mainly the intracellular ions K⁺ and Cl^–. Cell swelling–induced activation of separate K⁺ and Cl^– channels has been described in most preparations. Organic osmolytes are grouped in three categories: amino acids, polyalcohols, and methylamines. These osmolytes, particularly taurine, are present in …

Role Of Activator Protein-1 In The Down-Regulation Of The Human Cyp2j2 Gene In Hypoxia, Nicole Y. Marden, Eva Fiala-Beer, Shi-Hua Xiang, Michael Murray

School of Veterinary and Biomedical Sciences: Faculty Publications

The cytochrome P450 (CYP) 2J2 arachidonic acid epoxygenase gene was down-regulated at a pre-translational level in human hepatoma-derived HepG2 cells incubated in a hypoxic environment; under these conditions, the expression of c-Jun and c-Fos mRNA and protein was increased. The 5′-upstream region of the CYP2J2 gene was isolated by amplification of a 2341 bp fragment and putative regulatory elements that resembled activator protein-1 (AP-1)-like sequences were identified. From transient transfection analysis, c-Jun was found to strongly activate a CYP2J2–luciferase reporter construct, but co-transfection with plasmids encoding c-Fos or c-Fos-related antigens, Fra-1 and -2, abrogated reporter activity. Using a series …