Author Report for: Scott CNo contact information in database for Scott C
Eddleston M, Clutton RE, Taylor M, Thompson A, Worek F, John H, Thiermann H, Scott C Ref: Clinical Toxicology (Phila), :1, 2019 : PubMed ESTHER: Eddleston_2019_Clin.Toxicol.(Phila)__1 PubMedSearch: Eddleston 2019 Clin.Toxicol.(Phila) 1 PubMedID: 31452424 Abstract Objectives: Current therapeutic options for organophosphorus (OP) insecticide self-poisoning including atropine and oximes are inadequate and case fatality may exceed 20%. An OP hydrolase enzyme, OpdA, has been used for environmental cleansing of OP insecticides and prevented death in rat and non-human primate models of OP insecticide poisoning if given very quickly after exposure. We here tested OpdA's ability to break down OP insecticides in human serum and in clinically relevant minipig models of OP insecticide poisoning. Methods: Human serum was spiked with seven diverse WHO Class II OP insecticides (chlorpyrifos, quinalphos, diazinon, dimethoate, fenthion, phenthoate, and profenofos) and the effect of OpdA on degradation measured. The pharmacodynamic and clinical effects of OpdA treatment were studied in Gottingen minipigs orally poisoned with agricultural formulations of dimethoate EC40 or methyl parathion EC60; pharmacodynamic effects were also assessed in profenofos EC50-poisoned pigs. Results: OpdA effectively hydrolysed OP insecticides in human serum, with rates varying from 856 (SD 44) down to 0.107 (SD 0.01) moles of substrate hydrolysed/mole of enzyme/sec (kcat) for quinalphos and phenthoate, respectively, although at rates 2-3 log orders less than found in vitro in buffered solution. It showed clinical benefit in minipig models, reducing the dose of noradrenaline required to sustain an adequate mean arterial pressure after dimethoate (mean 0.149 [SD 0.10] mug/kg/h vs. 1.07 [SD 0.77] mug/kg/h, p < .0001) and methyl parathion (mean 0.077 [SD 0.08] mug/kg/h vs. 0.707 [SD 0.49] mug/kg/h, p < .0001) poisoning. OpdA reduced blood OP insecticide concentration and acetylcholinesterase inhibition after poisoning by dimethoate, methyl parathion, and profenofos insecticides. Conclusions: In vitro incubation of OpdA in human serum showed hydrolysis of diverse OP insecticides, although at lower rates than found in buffer solutions. This activity results in clinical and pharmacodynamic efficacy in vivo against several OP insecticides. These results support the testing of OpdA in further animal models before considering human trials to determine whether it may become an urgently required novel therapeutic agent for OP insecticide self-poisoning. Title: Evolutionary expansion of the amidohydrolase superfamily in bacteria in response to the synthetic compounds molinate and diuron Sugrue E, Fraser NJ, Hopkins DH, Carr PD, Khurana JL, Oakeshott JG, Scott C, Jackson CJ Ref: Applied Environmental Microbiology, 81:2612, 2015 : PubMed ESTHER: Sugrue_2015_Appl.Environ.Microbiol_81_2612 PubMedSearch: Sugrue 2015 Appl.Environ.Microbiol 81 2612 PubMedID: 25636851 Inhibitor(s) related to this paper: Diuron, Molinate Abstract The amidohydrolase superfamily has remarkable functional diversity, with considerable structural and functional annotation of known sequences. In microbes, the recent evolution of several members of this family to catalyze the breakdown of environmental xenobiotics is not well understood. An evolutionary transition from binuclear to mononuclear metal ion coordination at the active sites of these enzymes could produce large functional changes such as those observed in nature, but there are few clear examples available to support this hypothesis. To investigate the role of binuclear-mononuclear active-site transitions in the evolution of new function in this superfamily, we have characterized two recently evolved enzymes that catalyze the hydrolysis of the synthetic herbicides molinate (MolA) and phenylurea (PuhB). In this work, the crystal structures, mutagenesis, metal ion analysis, and enzyme kinetics of both MolA and PuhB establish that these enzymes utilize a mononuclear active site. However, bioinformatics and structural comparisons reveal that the closest putative ancestor of these enzymes had a binuclear active site, indicating that a binuclear-mononuclear transition has occurred. These proteins may represent examples of evolution modifying the characteristics of existing catalysts to satisfy new requirements, specifically, metal ion rearrangement leading to large leaps in activity that would not otherwise be possible. Title: Isomer-specific comparisons of the hydrolysis of synthetic pyrethroids and their fluorogenic analogues by esterases from the cotton bollworm Helicoverpa armigera Yuan G, Li Y, Farnsworth CA, Coppin CW, Devonshire AL, Scott C, Russell RJ, Wu Y, Oakeshott JG Ref: Pestic Biochem Physiol, 121:102, 2015 : PubMed ESTHER: Yuan_2015_Pestic.Biochem.Physiol_121_102 PubMedSearch: Yuan 2015 Pestic.Biochem.Physiol 121 102 PubMedID: 26047117 Abstract The low aqueous solubility and chiral complexity of synthetic pyrethroids, together with large differences between isomers in their insecticidal potency, have hindered the development of meaningful assays of their metabolism and metabolic resistance to them. To overcome these problems, Shan and Hammock (2001) [7] therefore developed fluorogenic and more water-soluble analogues of all the individual isomers of the commonly used Type 2 pyrethroids, cypermethrin and fenvalerate. The analogues have now been used in several studies of esterase-based metabolism and metabolic resistance. Here we test the validity of these analogues by quantitatively comparing their hydrolysis by a battery of 22 heterologously expressed insect esterases with the hydrolysis of the corresponding pyrethroid isomers by these esterases in an HPLC assay recently developed by Teese et al. (2013) [14]. We find a strong, albeit not complete, correlation (r = 0.7) between rates for the two sets of substrates. The three most potent isomers tested were all relatively slowly degraded in both sets of data but three esterases previously associated with pyrethroid resistance in Helicoverpa armigera did not show higher activities for these isomers than did allelic enzymes derived from susceptible H. armigera. Given their amenability to continuous assays at low substrate concentrations in microplate format, and ready detection of product, we endorse the ongoing utility of the analogues in many metabolic studies of pyrethroids. Title: A 5000-fold increase in the specificity of a bacterial phosphotriesterase for malathion through combinatorial active site mutagenesis Naqvi T, Warden AC, French N, Sugrue E, Carr PD, Jackson CJ, Scott C Ref: PLoS ONE, 9:e94177, 2014 : PubMed ESTHER: Naqvi_2014_PLoS.One_9_e94177 PubMedSearch: Naqvi 2014 PLoS.One 9 e94177 PubMedID: 24721933 Abstract Phosphotriesterases (PTEs) have been isolated from a range of bacterial species, including Agrobcaterium radiobacter (PTEAr), and are efficient enzymes with broad substrate ranges. The turnover rate of PTEAr for the common organophosphorous insecticide malathion is lower than expected based on its physical properties; principally the pka of its leaving group. In this study, we rationalise the turnover rate of PTEAr for malathion using computational docking of the substrate into a high resolution crystal structure of the enzyme, suggesting that malathion is too large for the PTEAr binding pocket. Protein engineering through combinatorial active site saturation testing (CASTing) was then used to increase the rate of malathion turnover. Variants from a CASTing library in which Ser308 and Tyr309 were mutated yielded variants with increased activity towards malathion. The most active PTEAr variant carried Ser308Leu and Tyr309Ala substitutions, which resulted in a ca. 5000-fold increase in kcat/KM for malathion. X-ray crystal structures for the PTEAr Ser308Leu\Tyr309Ala variant demonstrate that the access to the binding pocket was enhanced by the replacement of the bulky Tyr309 residue with the smaller alanine residue. Title: The zebrafish reference genome sequence and its relationship to the human genome Howe K, Clark MD, Torroja CF, Torrance J, Berthelot C, Muffato M, Collins JE, Humphray S, McLaren K and Stemple DL <167 more author(s)> Howe K, Clark MD, Torroja CF, Torrance J, Berthelot C, Muffato M, Collins JE, Humphray S, McLaren K, Matthews L, Mclaren S, Sealy I, Caccamo M, Churcher C, Scott C, Barrett JC, Koch R, Rauch GJ, White S, Chow W, Kilian B, Quintais LT, Guerra-Assuncao JA, Zhou Y, Gu Y, Yen J, Vogel JH, Eyre T, Redmond S, Banerjee R, Chi J, Fu B, Langley E, Maguire SF, Laird GK, Lloyd D, Kenyon E, Donaldson S, Sehra H, Almeida-King J, Loveland J, Trevanion S, Jones M, Quail M, Willey D, Hunt A, Burton J, Sims S, McLay K, Plumb B, Davis J, Clee C, Oliver K, Clark R, Riddle C, Elliot D, Threadgold G, Harden G, Ware D, Begum S, Mortimore B, Kerry G, Heath P, Phillimore B, Tracey A, Corby N, Dunn M, Johnson C, Wood J, Clark S, Pelan S, Griffiths G, Smith M, Glithero R, Howden P, Barker N, Lloyd C, Stevens C, Harley J, Holt K, Panagiotidis G, Lovell J, Beasley H, Henderson C, Gordon D, Auger K, Wright D, Collins J, Raisen C, Dyer L, Leung K, Robertson L, Ambridge K, Leongamornlert D, McGuire S, Gilderthorp R, Griffiths C, Manthravadi D, Nichol S, Barker G, Whitehead S, Kay M, Brown J, Murnane C, Gray E, Humphries M, Sycamore N, Barker D, Saunders D, Wallis J, Babbage A, Hammond S, Mashreghi-Mohammadi M, Barr L, Martin S, Wray P, Ellington A, Matthews N, Ellwood M, Woodmansey R, Clark G, Cooper J, Tromans A, Grafham D, Skuce C, Pandian R, Andrews R, Harrison E, Kimberley A, Garnett J, Fosker N, Hall R, Garner P, Kelly D, Bird C, Palmer S, Gehring I, Berger A, Dooley CM, Ersan-Urun Z, Eser C, Geiger H, Geisler M, Karotki L, Kirn A, Konantz J, Konantz M, Oberlander M, Rudolph-Geiger S, Teucke M, Lanz C, Raddatz G, Osoegawa K, Zhu B, Rapp A, Widaa S, Langford C, Yang F, Schuster SC, Carter NP, Harrow J, Ning Z, Herrero J, Searle SM, Enright A, Geisler R, Plasterk RH, Lee C, Westerfield M, de Jong PJ, Zon LI, Postlethwait JH, Nusslein-Volhard C, Hubbard TJ, Roest Crollius H, Rogers J, Stemple DL (- 167) Ref: Nature, 496:498, 2013 : PubMed ESTHER: Howe_2013_Nature_496_498 PubMedSearch: Howe 2013 Nature 496 498 PubMedID: 23594743 Gene_locus related to this paper: danre-1neur, danre-ABHD10b, danre-a9jrf7, danre-d2x2g3, danre-e7ezq9, danre-e7ff77, danre-ndr3, danre-nlgn4a, danre-q1mti5, danre-q6nyz4, danre-q6p2u2, danre-q7t359, danre-q08c93, danre-A2BGU9, danre-f1q676, danre-e7f0z8, danre-e7ez27, danre-e7f2w1, danre-f1qid7, danre-a0a0g2kru2, danre-f1qla7, danre-a9jr90, danre-e7f070, danre-f172a, danre-e7fb35, danre-a7mbu9, danre-f1qtr2 Abstract Zebrafish have become a popular organism for the study of vertebrate gene function. The virtually transparent embryos of this species, and the ability to accelerate genetic studies by gene knockdown or overexpression, have led to the widespread use of zebrafish in the detailed investigation of vertebrate gene function and increasingly, the study of human genetic disease. However, for effective modelling of human genetic disease it is important to understand the extent to which zebrafish genes and gene structures are related to orthologous human genes. To examine this, we generated a high-quality sequence assembly of the zebrafish genome, made up of an overlapping set of completely sequenced large-insert clones that were ordered and oriented using a high-resolution high-density meiotic map. Detailed automatic and manual annotation provides evidence of more than 26,000 protein-coding genes, the largest gene set of any vertebrate so far sequenced. Comparison to the human reference genome shows that approximately 70% of human genes have at least one obvious zebrafish orthologue. In addition, the high quality of this genome assembly provides a clearer understanding of key genomic features such as a unique repeat content, a scarcity of pseudogenes, an enrichment of zebrafish-specific genes on chromosome 4 and chromosomal regions that influence sex determination. Title: Organophosphate and pyrethroid hydrolase activities of mutant Esterases from the cotton bollworm Helicoverpa armigera Li Y, Farnsworth CA, Coppin CW, Teese MG, Liu JW, Scott C, Zhang X, Russell RJ, Oakeshott JG Ref: PLoS ONE, 8:e77685, 2013 : PubMed ESTHER: Li_2013_PLoS.One_8_e77685 PubMedSearch: Li 2013 PLoS.One 8 e77685 PubMedID: 24204917 Gene_locus related to this paper: helam-d5g3c9, helam-d5g3d2, helam-d5g3d3, helam-d5g3d4, helam-d5g3d5, helam-d5kx87, helam-d5kx99, helam-d5kxa9, helam-d9iv61, helam-d9iv62, helam-h9zvh4, helam-s4wfz6 Abstract Two mutations have been found in five closely related insect esterases (from four higher Diptera and a hymenopteran) which each confer organophosphate (OP) hydrolase activity on the enzyme and OP resistance on the insect. One mutation converts a Glycine to an Aspartate, and the other converts a Tryptophan to a Leucine in the enzymes' active site. One of the dipteran enzymes with the Leucine mutation also shows enhanced activity against pyrethroids. Introduction of the two mutations in vitro into eight esterases from six other widely separated insect groups has also been reported to increase substantially the OP hydrolase activity of most of them. These data suggest that the two mutations could contribute to OP, and possibly pyrethroid, resistance in a variety of insects. We therefore introduced them in vitro into eight Helicoverpa armigera esterases from a clade that has already been implicated in OP and pyrethroid resistance. We found that they do not generally enhance either OP or pyrethroid hydrolysis in these esterases but the Aspartate mutation did increase OP hydrolysis in one enzyme by about 14 fold and the Leucine mutation caused a 4-6 fold increase in activity (more in one case) of another three against some of the most insecticidal isomers of fenvalerate and cypermethrin. The Aspartate enzyme and one of the Leucine enzymes occur in regions of the H. armigera esterase isozyme profile that have been previously implicated in OP and pyrethroid resistance, respectively. Title: How many genetic options for evolving insecticide resistance in heliothine and spodopteran pests? Oakeshott JG, Farnsworth CA, East PD, Scott C, Han Y, Wu Y, Russell RJ Ref: Pest Manag Sci, 69:889, 2013 : PubMed ESTHER: Oakeshott_2013_Pest.Manag.Sci_69_889 PubMedSearch: Oakeshott 2013 Pest.Manag.Sci 69 889 PubMedID: 23526801 Abstract The widely accepted paradigm for the development of insecticide resistance in field populations of insects is of selection for one or a very few genes of major effect. Limited genetic mapping data for organophosphate and pyrethroid resistance in heliothine and spodopteran pests generally agrees with this paradigm. However, other biochemical and transcriptomic data suggest a more complex set of changes in multiple P450 and esterase gene/enzyme systems in resistant strains of these species. We discuss possible explanations for this paradox, including the likely embedding of these genes in regulatory cascades and emerging evidence for their arrangement in large clusters of closely related genes. We conclude that there could indeed be an unusually large number of genetic options for evolving resistance in these species. Title: Heterologous expression and biochemical characterisation of fourteen esterases from Helicoverpa armigera Teese MG, Farnsworth CA, Li Y, Coppin CW, Devonshire AL, Scott C, East P, Russell RJ, Oakeshott JG Ref: PLoS ONE, 8:e65951, 2013 : PubMed ESTHER: Teese_2013_PLoS.ONE_8_E65951 PubMedSearch: Teese 2013 PLoS.ONE 8 E65951 PubMedID: 23799064 Gene_locus related to this paper: helam-d5g3c9, helam-s4wfz6 Abstract Esterases have recurrently been implicated in insecticide resistance in Helicoverpa armigera but little is known about the underlying molecular mechanisms. We used a baculovirus system to express 14 of 30 full-length esterase genes so far identified from midgut cDNA libraries of this species. All 14 produced esterase isozymes after native PAGE and the isozymes for seven of them migrated to two regions of the gel previously associated with both organophosphate and pyrethroid resistance in various strains. Thirteen of the enzymes obtained in sufficient yield for further analysis all showed tight binding to organophosphates and low but measurable organophosphate hydrolase activity. However there was no clear difference in activity between the isozymes from regions associated with resistance and those from elsewhere in the zymogram, or between eight of the isozymes from a phylogenetic clade previously associated with resistance in proteomic and quantitative rtPCR experiments and five others not so associated. By contrast, the enzymes differed markedly in their activities against nine pyrethroid isomers and the enzymes with highest activity for the most insecticidal isomers were from regions of the gel and, in some cases, the phylogeny that had previously been associated with pyrethroid resistance. Phospholipase treatment confirmed predictions from sequence analysis that three of the isozymes were GPI anchored. This unusual feature among carboxylesterases has previously been suggested to underpin an association that some authors have noted between esterases and resistance to the Cry1Ac toxin from Bacillus thuringiensis. However these three isozymes did not migrate to the zymogram region previously associated with Cry1Ac resistance. Title: Esterase-based metabolic resistance to insecticides in heliothine and spodopteran pests Farnsworth CA, Teese MG, Yuan G, Li Y, Scott C, Zhang X, Wu Y, Russell RJ, Oakeshott JG Ref: Journal of Pesticide Science, 35:275, 2010 : PubMed ESTHER: Farnsworth_2010_J.Pestic.Sci_35_275 PubMedSearch: Farnsworth 2010 J.Pestic.Sci 35 275 Abstract Elevated esterase activities and increased band intensities of multiple esterase isozymes after electrophoresis are commonly associated with resistance to organophosphate, pyrethroid and carbamate insecticides in various heliothine and spodopteran pests. One possible explanation for this involves a "master regulator" mutation in a more general chemical stress response. An association between elevated esterase activities and isozyme intensities has also been reported for resistance to the Cry1Ac toxin of Helicoverpa armigera. The basis for this is unclear albeit some involvement of esterases could be mediated by the toxin's affinity for N-acetyl galactosamine glycans on certain gut-expressed esterases in this species. Title: Pharmacokinetics of OpdA, an organophosphorus hydrolase, in the African green monkey Jackson CJ, Scott C, Carville A, Mansfield K, Ollis DL, Bird SB Ref: Biochemical Pharmacology, 80:1075, 2010 : PubMed ESTHER: Jackson_2010_Biochem.Pharmacol_80_1075 PubMedSearch: Jackson 2010 Biochem.Pharmacol 80 1075 PubMedID: 20599794 Abstract Organophosphorus (OP) pesticides are a broad class of acetylcholinesterase inhibitors that are responsible for tremendous morbidity and mortality worldwide, contributing to an estimated 300,000 deaths annually. Current pharmacotherapy for acute OP poisoning includes the use of atropine, an oxime, and benzodiazepines. However, even with such therapy, the mortality from these agents are as high as 40%. Enzymatic hydrolysis of OPs is an attractive new potential therapy for acute OP poisoning. A number of bacterial OP hydrolases have been isolated. A promising OP hydrolase is an enzyme isolated from Agrobacterium radiobacter, named OpdA. OpdA has been shown to decrease lethality in rodent models of parathion and dichlorvos poisoning. However, pharmacokinetic data have not been obtained. In this study, we examined the pharmacokinetics of OpdA in an African Green Monkey model. At a dose of 1.2mg/kg the half-life of OpdA was approximately 40 min, with a mean residence time of 57 min. As expected, the half-life did not change with the dose of OpdA given: at doses of 0.15 and 0.45 mg/kg, the half-life of OpdA was 43.1 and 38.9 min, respectively. In animals subjected to 5 daily doses of OpdA, the residual activity that was measured 24h after each OpdA dose increased 5-fold for the 0.45 mg/kg dose and 11-fold for the 1.2mg/kg dose. OpdA exhibits pharmacokinetics favorable for the further development as a therapy for acute OP poisoning, particularly for hydrophilic OP pesticides. Future work to increase the half-life of OpdA may be beneficial. Title: Gene identification and proteomic analysis of the esterases of the cotton bollworm, Helicoverpa armigera Teese MG, Campbell PM, Scott C, Gordon KH, Southon A, Hovan D, Robin C, Russell RJ, Oakeshott JG Ref: Insect Biochemistry & Molecular Biology, 40:1, 2010 : PubMed ESTHER: Teese_2010_Insect.Biochem.Mol.Biol_40_1 PubMedSearch: Teese 2010 Insect.Biochem.Mol.Biol 40 1 PubMedID: 20005949 Gene_locus related to this paper: helam-d5g3c9, helam-d5g3d2, helam-d5g3d3, helam-d5g3d4, helam-d5g3d5, helam-d5g3d9, helam-d5g3e0, helam-d5g3e1, helam-d5g3e3, helam-d5g3e4, helam-d5g3e6, helam-d5g3e7, helam-d5g3e8, helam-d5g3f1, helam-d5g3f2, helam-d5g3f3, helam-d5g3f4, helam-d5g3f9, helam-d5g3g0, helam-d5g3g1, helam-d5g3g3, helam-d5kx99, helam-d5kxa9, helam-d9iv61, helam-f8rgs7, helam-h9zvh4 Abstract Some of the resistance of Helicoverpa armigera to conventional insecticides such as organophosphates and synthetic pyrethroids appears to be due to metabolic detoxification by carboxylesterases. To investigate the H. armigera carboxyl/cholinesterases, we created a data set of 39 putative paralogous H. armigera carboxyl/cholinesterase sequences from cDNA libraries and other sources. Phylogenetic analysis revealed a close relationship between these sequences and 70 carboxyl/cholinesterases from the recently sequenced genome of the silkworm, Bombyx mori, including several conserved clades of non-catalytic proteins. A juvenile hormone esterase candidate from H. armigera was identified, and B. mori orthologues were proposed for 31% of the sequences examined, however low similarity was found between lepidopteran sequences and esterases previously associated with insecticide resistance from other insect orders. A proteomic analysis of larval esterases then enabled us to match seven of the H. armigera carboxyl/cholinesterase sequences to specific esterase isozymes. All identified sequences were predicted to encode catalytically active carboxylesterases, including six proteins with N-terminal signal peptides and N-glycans, with two also containing C-terminal signals for glycosylphosphatidylinositol anchor attachment. Five of these sequences were matched to zones of activity on native PAGE at relative mobility values previously associated with insecticide resistance in this species. Title: Characterization of the phenylurea hydrolases A and B: founding members of a novel amidohydrolase subgroup Khurana JL, Jackson CJ, Scott C, Pandey G, Horne I, Russell RJ, Herlt A, Easton CJ, Oakeshott JG Ref: Biochemical Journal, 418:431, 2009 : PubMed ESTHER: Khurana_2009_Biochem.J_418_431 PubMedSearch: Khurana 2009 Biochem.J 418 431 PubMedID: 19000034 Gene_locus related to this paper: 9myco-b8r4k2 Abstract Mycobacterium brisbanense strain JK1, a bacterium capable of degrading the herbicide diuron, was isolated from herbicide-exposed soil. A gene/enzyme system with diuron hydrolase activity was isolated from this strain and named PUH (phenylurea hydrolase) B (puhB/PuhB) because of its close similarity to the previously characterized PUH A (puhA/PuhA). Both PUHs were heterologously expressed, purified and characterized. The PUHs were found to oligomerize as hexamers in solution, with each monomer containing a mononuclear Zn2+ active site. Sequence analysis showed that these enzymes belong to the metal-dependent amidohydrolase superfamily, although they contain a hitherto unreported Asn-X-His metal-binding motif and appear to form a novel sub-group within this superfamily. The effects of temperature and solvent on the enzymes were characterized. Determination of the kinetic parameters of the PUHs was used alongside Bronsted plots to develop a plausible catalytic mechanism, which is similar to that used by urease. In addition to the primary PUH activity, both enzymes are catalytically promiscuous, efficiently hydrolysing esters, carbamates and phosphotriesters. In fact, an analogue of diuron, in which the C-N bond was replaced by a C-O bond, was found to be turned over as efficiently as diuron, suggesting that the substrate specificity is predominantly determined by steric factors. The discovery of PuhA and PuhB on separate continents, and the absence of any other close homologues in the available sequence databases, poses a challenging question regarding the evolutionary origins of these enzymes. Title: Heterologous expression of the methyl carbamate-degrading hydrolase MCD Naqvi T, Cheesman MJ, Williams MR, Campbell PM, Ahmed S, Russell RJ, Scott C, Oakeshott JG Ref: J Biotechnol, 144:89, 2009 : PubMed ESTHER: Naqvi_2009_J.Biotechnol_144_89 PubMedSearch: Naqvi 2009 J.Biotechnol 144 89 PubMedID: 19770008 Abstract The methyl carbamate-degrading hydrolase (MCD) of Achromobacter WM111 has considerable potential as a pesticide bioremediation agent. However this potential has been unrealisable until now because of an inability to express MCD in heterologous hosts such as Escherichia coli. Herein, we describe the first successful attempt to express appreciable quantities of MCD in active form in E. coli, and the subsequent characterisation of the heterologously expressed material. We find that the properties of this material closely match the previously reported properties of MCD produced from Achromobacter WM111. This includes the presence of two distinct forms of the enzyme that we show are most likely due to the presence of two functional translational start sites. The purified enzyme catalyses the hydrolysis of a carbamate (carbaryl), a carboxyl ester (alpha-naphthyl acetate) and a phophotriester (dimethyl umbelliferyl phosphate) and it is relatively resistant to thermal and solvent-mediated denaturation. The robust nature and catalytic promiscuity of MCD suggest that it could be exploited for various biotechnological applications. Title: The DNA sequence and comparative analysis of human chromosome 10 Deloukas P, Earthrowl ME, Grafham DV, Rubenfield M, French L, Steward CA, Sims SK, Jones MC, Searle S and Rogers J <126 more author(s)> Deloukas P, Earthrowl ME, Grafham DV, Rubenfield M, French L, Steward CA, Sims SK, Jones MC, Searle S, Scott C, Howe K, Hunt SE, Andrews TD, Gilbert JG, Swarbreck D, Ashurst JL, Taylor A, Battles J, Bird CP, Ainscough R, Almeida JP, Ashwell RI, Ambrose KD, Babbage AK, Bagguley CL, Bailey J, Banerjee R, Bates K, Beasley H, Bray-Allen S, Brown AJ, Brown JY, Burford DC, Burrill W, Burton J, Cahill P, Camire D, Carter NP, Chapman JC, Clark SY, Clarke G, Clee CM, Clegg S, Corby N, Coulson A, Dhami P, Dutta I, Dunn M, Faulkner L, Frankish A, Frankland JA, Garner P, Garnett J, Gribble S, Griffiths C, Grocock R, Gustafson E, Hammond S, Harley JL, Hart E, Heath PD, Ho TP, Hopkins B, Horne J, Howden PJ, Huckle E, Hynds C, Johnson C, Johnson D, Kana A, Kay M, Kimberley AM, Kershaw JK, Kokkinaki M, Laird GK, Lawlor S, Lee HM, Leongamornlert DA, Laird G, Lloyd C, Lloyd DM, Loveland J, Lovell J, Mclaren S, McLay KE, McMurray A, Mashreghi-Mohammadi M, Matthews L, Milne S, Nickerson T, Nguyen M, Overton-Larty E, Palmer SA, Pearce AV, Peck AI, Pelan S, Phillimore B, Porter K, Rice CM, Rogosin A, Ross MT, Sarafidou T, Sehra HK, Shownkeen R, Skuce CD, Smith M, Standring L, Sycamore N, Tester J, Thorpe A, Torcasso W, Tracey A, Tromans A, Tsolas J, Wall M, Walsh J, Wang H, Weinstock K, West AP, Willey DL, Whitehead SL, Wilming L, Wray PW, Young L, Chen Y, Lovering RC, Moschonas NK, Siebert R, Fechtel K, Bentley D, Durbin R, Hubbard T, Doucette-Stamm L, Beck S, Smith DR, Rogers J (- 126) Ref: Nature, 429:375, 2004 : PubMed ESTHER: Deloukas_2004_Nature_429_375 PubMedSearch: Deloukas 2004 Nature 429 375 PubMedID: 15164054 Gene_locus related to this paper: human-LIPA, human-LIPK, human-PNLIPRP1, human-PNLIPRP2, human-PNLIPRP3 Abstract The finished sequence of human chromosome 10 comprises a total of 131,666,441 base pairs. It represents 99.4% of the euchromatic DNA and includes one megabase of heterochromatic sequence within the pericentromeric region of the short and long arm of the chromosome. Sequence annotation revealed 1,357 genes, of which 816 are protein coding, and 430 are pseudogenes. We observed widespread occurrence of overlapping coding genes (either strand) and identified 67 antisense transcripts. Our analysis suggests that both inter- and intrachromosomal segmental duplications have impacted on the gene count on chromosome 10. Multispecies comparative analysis indicated that we can readily annotate the protein-coding genes with current resources. We estimate that over 95% of all coding exons were identified in this study. Assessment of single base changes between the human chromosome 10 and chimpanzee sequence revealed nonsense mutations in only 21 coding genes with respect to the human sequence. Title: Genome sequence of the plant pathogen and biotechnology agent Agrobacterium tumefaciens C58 Goodner B, Hinkle G, Gattung S, Miller N, Blanchard M, Qurollo B, Goldman BS, Cao Y, Askenazi M and Slater S <21 more author(s)> Goodner B, Hinkle G, Gattung S, Miller N, Blanchard M, Qurollo B, Goldman BS, Cao Y, Askenazi M, Halling C, Mullin L, Houmiel K, Gordon J, Vaudin M, Iartchouk O, Epp A, Liu F, Wollam C, Allinger M, Doughty D, Scott C, Lappas C, Markelz B, Flanagan C, Crowell C, Gurson J, Lomo C, Sear C, Strub G, Cielo C, Slater S (- 21) Ref: Science, 294:2323, 2001 : PubMed ESTHER: Goodner_2001_Science_294_2323 PubMedSearch: Goodner 2001 Science 294 2323 PubMedID: 11743194 Gene_locus related to this paper: agrt5-a9cf94, agrt5-a9cfa9, agrt5-a9cfs8, agrt5-a9cfu7, agrt5-a9cie7, agrt5-a9cj11, agrt5-a9cjp2, agrt5-a9cki2, agrt5-a9ckr2, agrt5-a9ckt2, agrt5-a9cle4, agrt5-a9clq8, agrt5-a9clq9, agrt5-q7cx24, agrt5-q7d1j0, agrt5-q7d1j3, agrt5-q7d3m5, agrt5-q7d3t6, agrt5-y5261, agrtu-ACVB, agrtu-ATTS, agrtu-ATU0253, agrtu-ATU0403, agrtu-ATU0841, agrtu-ATU1045, agrtu-ATU1102, agrtu-ATU1572, agrtu-ATU1617, agrtu-ATU1826, agrtu-ATU1842, agrtu-ATU2061, agrtu-ATU2126, agrtu-ATU2171, agrtu-ATU2409, agrtu-ATU2452, agrtu-ATU2481, agrtu-ATU2497, agrtu-ATU2576, agrtu-ATU3428, agrtu-ATU3651, agrtu-ATU3652, agrtu-ATU4238, agrtu-ATU5190, agrtu-ATU5193, agrtu-ATU5275, agrtu-ATU5296, agrtu-ATU5348, agrtu-ATU5389, agrtu-ATU5446, agrtu-ATU5495, agrtu-CPO, agrtu-DHAA, agrtu-DLHH, agrtu-EPHA, agrtu-GRST, agrtu-PCA, agrtu-PCAD, agrtu-PHBC, agrtu-PTRB, agrt5-a9cji8 Abstract Agrobacterium tumefaciens is a plant pathogen capable of transferring a defined segment of DNA to a host plant, generating a gall tumor. Replacing the transferred tumor-inducing genes with exogenous DNA allows the introduction of any desired gene into the plant. Thus, A. tumefaciens has been critical for the development of modern plant genetics and agricultural biotechnology. Here we describe the genome of A. tumefaciens strain C58, which has an unusual structure consisting of one circular and one linear chromosome. We discuss genome architecture and evolution and additional genes potentially involved in virulence and metabolic parasitism of host plants. Title: Genetic and Morphological Characterization of Cladobotryum Species Causing Cobweb Disease of Mushrooms McKay GJ, Egan D, Morris E, Scott C, Brown AE Ref: Applied Environmental Microbiology, 65:606, 1999 : PubMed ESTHER: McKay_1999_Appl.Environ.Microbiol_65_606 PubMedSearch: McKay 1999 Appl.Environ.Microbiol 65 606 PubMedID: 9925589 Abstract Cladobotryum dendroides (= Dactylium dendroides) has hitherto been regarded as the major causal agent of cobweb disease of the cultivated mushroom, Agaricus bisporus. Nucleotide sequence data for the internal transcribed spacer (ITS) regions of four Cladobotryum/Hypomyces species reported to be associated with cobweb disease, however, indicate that the most common pathogen is now C. mycophilum. This cobweb pathogen varies somewhat in conidial septation from published descriptions of C. mycophilum and lacks the distinctive colony odor. ITS sequencing revealed minor nucleotide variation which split isolates of the pathogen into three subgroups, two comprising isolates that were sensitive to methylbenzimidazole carbamate (MBC) fungicides and one comprising MBC-resistant isolates. The MBC-resistant isolates, which were only obtained from Ireland and Great Britain, clustered together strongly in randomly amplified polymorphic DNA (RAPD) PCR analysis, suggesting that they may be clonal. The MBC-sensitive isolates were more diverse. A RAPD fragment of 800 to 900 bp, containing a microsatellite and found in the MBC-resistant isolates, also indicated their clonal nature; the microsatellites of these isolates contained the same number of GA repeats. Smaller, polymorphic microsatellites, similarly comprising GA repeats, in the MBC-sensitive isolates in general correlated with their geographic origin. | |||||||
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