“Out of the Can”: A Draft Genome Assembly, Liver Transcriptome, and Nutrigenomics of the European Sardine, Sardina pilchardus

Clupeiformes, such as sardines and herrings, represent an important share of worldwide fisheries. Among those, the European sardine (Sardina pilchardus, Walbaum 1792) exhibits significant commercial relevance. While the last decade showed a steady and sharp decline in capture levels, recent advances in culture husbandry represent promising research avenues. Yet, the complete absence of genomic resources from sardine imposes a severe bottleneck to understand its physiological and ecological requirements. We generated 69 Gbp of paired-end reads using Illumina HiSeq X Ten and assembled a draft genome assembly with an N50 scaffold length of 25,579 bp and BUSCO completeness of 82.1% (Actinopterygii). The estimated size of the genome ranges between 655 and 850 Mb. Additionally, we generated a relatively high-level liver transcriptome. To deliver a proof of principle of the value of this dataset, we established the presence and function of enzymes (Elovl2, Elovl5, and Fads2) that have pivotal roles in the biosynthesis of long chain polyunsaturated fatty acids, essential nutrients particularly abundant in oily fish such as sardines. Our study provides the first sustainableomics datasetexploitation.from a valuable economic marine teleost species, the European sardine, representing an essential resource for their effective conservation, management, and sustainable exploitation. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.Funding: We acknowledge the North Portugal Regional Operational Program (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) that supported this research through the Coral—Sustainable Ocean Exploitation (reference NORTE-01-0145-FEDER-000036). R.R.d.F. thanks the Danish National Research Foundation for its support of the Center for Macroecology, Evolution, and Climate (grant DNRF96). Acknowledgments: Some computational work was performed on the Abel Supercomputing Cluster (Norwegian metacenter for High Performance Computing (NOTUR) and the University of Oslo) operated by the Research Computing Services group at USIT, the University of Oslo IT-department (http://www.hpc.uio.no/). We would like to thank Jette Bornholdt, Amal Al-Chaer and George Pacheco for help with laboratory procedures, and the Bioinformatics Center of the University of Copenhagen for providing laboratory space. This work is part of the CIIMAR-lead initiative Portugal-Fishomics

Modulatory Effects Of Yerba Maté (ilex Paraguariensis) On The Pi3k-akt Signaling Pathway

The aim of this study was to evaluate the effects of yerba maté (YM) extract on the phosphatidylinositol 3-kinase (PI3K)-AKT signaling pathway in vivo. The mice were introduced to either standard- or high-fat diet (HFD). After 8 weeks on an HFD, mice were randomly assigned to one of the two treatment conditions, water or yerba maté extract at 1.0 g/kg. After treatment, glucose blood level and hepatic insulin response were evaluated. Liver tissue was examined to determine the mRNA levels using the PI3K-AKT PCR array. The nuclear translocation of forkhead box O1 (FOXO1) was determined by an electrophoretic mobility-shift assay. Our data demonstrated that yerba maté extract significantly decreased the final body weight, glucose blood levels, and insulin resistance of mice. Molecular analysis demonstrated that an HFD downregulated Akt2, Irs1, Irs2, Pi3kca, Pi3kcg, and Pdk1; after yerba maté treatment, the levels of those genes returned to baseline. In addition, an HFD upregulated Pepck and G6pc and increased FOXO1 nuclear translocation. The intervention downregulated these genes by decreasing FOXO1 nuclear translocation. The results obtained demonstrate for the first time the specific action of yerba maté on the PI3K-AKT pathway, which contributed to the observed improvement in hepatic insulin signaling. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.571018821885Ahima, R.S., Flier, J.S., Adipose tissue as an endocrine organ (2000) Trends Endocrinol. Metab., 11, pp. 327-332Ahima, R.S., Digging deeper into obesity (2011) J. Clin. Invest., 121, pp. 2076-2079Fruhbeck, G., Gomez-Ambrosi, J., Muruzabal, F.J., Burrell, M.A., The adipocyte: a model for integration of endocrine and metabolic signaling in energy metabolism regulation (2001) Am. J. Physiol. Endocrinol. Metab., 280, pp. E827-E847Lazar, M.A., How obesity causes diabetes: not a tall tale (2005) Science, 307, pp. 373-375Eckel, R.H., Grundy, S.M., Zimmet, P.Z., The metabolic syndrome (2005) Lancet, 365, pp. 1415-1428Taniguchi, C.M., Emanuelli, B., Kahn, C.R., Critical nodes in signalling pathways: insights into insulin action (2006) Nat. Rev. Mol. Cell Biol., 7, pp. 85-96Gonzalez-Yanes, C., Serrano, A., Bermudez-Silva, F.J., Hernandez-Dominguez, M., Oleylethanolamide impairs glucose tolerance and inhibits insulin-stimulated glucose uptake in rat adipocytes through p38 and JNK MAPK pathways (2005) Am. J. Physiol. Endocrinol. Metab., 289, pp. E923-E929Huang, T.H., Kota, B.P., Razmovski, V., Roufogalis, B.D., Herbal or natural medicines as modulators of peroxisome proliferator-activated receptors and related nuclear receptors for therapy of metabolic syndrome (2005) Basic Clin. Pharmacol. Toxicol., 96, pp. 3-14Hwang, J.T., Kwon, D.Y., Yoon, S.H., AMP-activated protein kinase: a potential target for the diseases prevention by natural occurring polyphenols (2009) New Biotechnol., 26, pp. 17-22Pang, J., Choi, Y., Park, T., Ilex paraguariensis extract ameliorates obesity induced by high-fat diet: potential role of AMPK in the visceral adipose tissue (2008) Arch. Biochem. Biophys., 476, pp. 178-185Arcari, D.P., Bartchewsky, W., Dos Santos, T.W., Oliveira, K.A., Antiobesity effects of yerba mate extract (Ilex paraguariensis) in high-fat diet-induced obese mice (2009) Obesity (Silver Spring), 17, pp. 2127-2133Arcari, D.P., Bartchewsky, W., Dos Santos, T.W., Oliveira, K.A., Anti-inflammatory effects of yerba mate extract (Ilex paraguariensis) ameliorate insulin resistance in mice with high fat diet-induced obesity (2011) Mol. Cell Endocrinol., 335, pp. 110-115Hussein, G.M., Matsuda, H., Nakamura, S., Hamao, M., Mate tea (Ilex paraguariensis) promotes satiety and body weight lowering in mice: involvement of glucagon-like peptide-1 (2011) Biol. Pharm. Bull., 34, pp. 1849-1855Kang, Y.R., Lee, H.Y., Kim, J.H., Moon, D.I., Anti-obesity and anti-diabetic effects of Yerba Mate (Ilex paraguariensis) in C57BL/6J mice fed a high-fat diet (2012) Lab. Anim. Res., 28, pp. 23-29Pimentel, G.D., Lira, F.S., Rosa, J.C., Caris, A.V., Yerba mate extract (Ilex paraguariensis) attenuates both central and peripheral inflammatory effects of diet-induced obesity in rats (2012) J. Nutr. Biochem., , DOI:10.1016/j.jnutbio.2012.04.016Gosmann, G., Barlette, A.G., Dhamer, T., Arcari, D.P., Phenolic compounds from mate (Ilex paraguariensis) inhibit adipogenesis in 3T3-L1 preadipocytes (2012) Plant Foods Hum. Nutr., 67, pp. 156-161Hussein, G.M., Matsuda, H., Nakamura, S., Akiyama, T., Protective and ameliorative effects of mate (Ilex paraguariensis) on metabolic syndrome in TSOD mice (2011) Phytomedicine, 19, pp. 88-97Mosimann, A.L., Wilhelm-Filho, D., Da Silva, E.L., Aqueous extract of Ilex paraguariensis attenuates the progression of atherosclerosis in cholesterol-fed rabbits (2006) Biofactors, 26, pp. 59-70Taha, C., Klip, A., The insulin signaling pathway (1999) J. Membr. Biol., 169, pp. 1-12White, M.F., IRS proteins and the common path to diabetes (2002) Am. J. Physiol. Endocrinol. Metab., 283, pp. E413-E422Previs, S.F., Withers, D.J., Ren, J.M., White, M.F.e.a., Contrasting effects of IRS-1 versus IRS-2 gene disruption on carbohydrate and lipid metabolism in vivo (2000) J. Biol. Chem., 275, pp. 38990-38994Whitehead, J.P., Humphreys, P., Krook, A., Jackson, R., Molecular scanning of the insulin receptor substrate 1 gene in subjects with severe insulin resistance: detection and functional analysis of a naturally occurring mutation in a YMXM motif (1998) Diabetes, 47, pp. 837-839Farese, R.V., Sajan, M.P., Standaert, M.L., Insulin-sensitive protein kinases (atypical protein kinase C and protein kinase B/Akt): actions and defects in obesity and type II diabetes (2005) Exp. Biol. Med. (Maywood), 230, pp. 593-605Michael, M.D., Kulkarni, R.N., Postic, C., Previs, S.F., Loss of insulin signaling in hepatocytes leads to severe insulin resistance and progressive hepatic dysfunction (2000) Mol. Cell, 6, pp. 87-97Mora, A., Lipina, C., Tronche, F., Sutherland, C., Deficiency of PDK1 in liver results in glucose intolerance, impairment of insulin-regulated gene expression and liver failure (2005) Biochem. J., 385, pp. 639-648Okamoto, Y., Ogawa, W., Nishizawa, A., Inoue, H., Restoration of glucokinase expression in the liver normalizes postprandial glucose disposal in mice with hepatic deficiency of PDK1 (2007) Diabetes, 56, pp. 1000-1009Chan, T.O., Tsichlis, P.N., PDK2: a complex tail in one Akt (2001) Sci STKE, 2001, pp. pe1Matsumoto, M., Han, S., Kitamura, T., Accili, D., Dual role of transcription factor FoxO1 in controlling hepatic insulin sensitivity and lipid metabolism (2006) J. Clin. Invest., 116, pp. 2464-2472Postic, C., Dentin, R., Girard, J., Role of the liver in the control of carbohydrate and lipid homeostasis (2004) Diabetes Metab., 30, pp. 398-40

Detection Of Gly-196-ser Mutation In 5α-reductase Type Ii Gene In A Brazilian Patient With Female Assignment And Behavior

We describe the identification of a single base mutation in the 5α- reductase type II gene in a Brazilian patient who was reared as female and remained with female behavior and sexual identity.113465466Labrie, F., Sugimoto, Y., Luu-The, V., Simard, J., Lachance, Y., Bachvarov, D., Leblanc, G., Paquet, N., Structure of human type II 5α-reductase gene (1992) Endocrinology, 131, pp. 1571-1573Russel, D.W., Wilson, J.D., Steroid 5α-reductase: Two genes/two enzymes (1994) Ann Rev Biochem, 63, pp. 25-61Thigpen, A.E., Davis, D.L., Miltovich, A., Mendonça, B.B., Imperato-McGinley, J., Griffin, J.E., Francke, U., Russel, D.W., Molecular genetics of 5α-reductase deficiency (1992) J Clin Invest, 90, pp. 799-809Katz, M.D., Cai, L.Q., Zhu, Y., Herrera, C., DeFillo-Ricart, M., Shackleton, H.L., Imperato-McGinley, J., The biochemical and phenotypic characterization of females homozygous for 5α-reductase deficiency (1995) J Clin Endocrinol Metab, 80, pp. 3160-3167Hiort, O., Willengring, H., Albers, N., Hecker, W., Engert, J., Dibbelt, L., Sinnecker, G.H.G., Molecular genetic analysis and human chorionic gonadotiopin stimulation tests in the diagnosis of prepubertal patients with partial 5α-reductase deficiency (1996) Eur J Pediatr, 155, pp. 445-451Sinnecker, G.H.G., Hiort, O., Dibbelt, L., Albers, N., Dörr, H.G., Hauss, H., Heinrich, U., Kruse, K., Phenotypic classification of male pseudohermaphroditism due to steroid 5α-reductase 2 deficiency (1996) Am J Med Genet, 63, pp. 223-23

Ribb And Ribba Genes From Acidithiobacillus Ferrooxidans: Expression Levels Under Different Growth Conditions And Phylogenetic Analysis

Acidithiobacillus ferrooxidans is a Gram-negative, chemolithoautotrophic bacterium involved in metal bioleaching. Using the RNA arbitrarily primed polymerase chain reaction (RAP-PCR), we have identified several cDNAs that were differentially expressed when A. ferrooxidans LR was submitted to potassium- and phosphate-limiting conditions. One of these cDNAs showed similarity with ribB. An analysis of the A. ferrooxidans ATCC 23270 genome, made available by The Institute for Genomic Research, showed that the ribB gene was not located in the rib operon, but a ribBA gene was present in this operon instead. The ribBA gene was isolated from A. ferrooxidans LR and expression of both ribB and ribBA was investigated. Transcript levels of both genes were enhanced in cells grown in the absence of K 2HPO 4, in the presence of zinc and copper sulfate and in different pHs. Transcript levels decreased upon exposure to a temperature higher than the ideal 30 °C and at pH 1.2. A comparative genomic analysis using the A. ferrooxidans ATCC 23270 genome revealed similar putative regulatory elements for both genes. Moreover, an RFN element was identified upstream from the ribB gene. Phylogenetic analysis of the distribution of RibB and RibBA in bacteria showed six different combinations. We suggest that the presence of duplicated riboflavin synthesis genes in bacteria must provide their host with some benefit in certain stressful situations. © 2008 Elsevier Masson SAS. All rights reserved.1596423431Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., Gapped BLAST and PSI-BLAST: a new generation of protein database search programs (1997) Nucleic Acids Research, 25, pp. 3389-3402Alvarez, S., Jerez, C.A., Copper ions stimulate polyphosphate degradation and phosphate efflux in Acidithiobacillus ferrooxidans (2004) Applied and Environmental Microbiology, 70, pp. 5177-5182Bacher, A., Eberhardt, S., Richter, G., Biosynthesis of riboflavin (1996) Escherichia coli and Salmonella typhimurium, Cellular and molecular biology, pp. 657-664. , Neidhart F.C., Curtiss III R., Ingraham J.L., Lin E.C.C., Low K.B., Magasanik B., Reznikoff W.S., Riley M., Schaechter M., and Umbarger H.E. (Eds), American Society for Microbiology, Washington, DCBateman, A., Coin, L., Durbin, R., Finn, R.D., Hollich, V., Griffiths-Jones, S., The Pfam protein families database (2004) Nucleic Acids Research, 32, pp. D138-D141Bereswill, S., Fassbinder, F., Volzing, C., Covacci, A., Haas, R., Kist, M., Hemolytic properties and riboflavin synthesis of Helicobacter pylori: cloning and functional characterization of the ribA gene encoding GTP-cyclohydrolase II that confers hemolytic activity to Escherichia coli (1998) Medical Microbiology and Immunology, 186, pp. 177-187Bosecker, K., Bioleaching: metal solubilization by microorganisms (1997) FEMS Microbiology Reviews, 20, pp. 591-604Clamp, M., Cuff, J., Searle, S.M., Barton, G.J., The Jalview Java alignment editor (2004) Bioinformatics, 20, pp. 426-427Cobley, J.G., Cox, J.C., Energy conservation in acidophilic bacteria (1983) Microbiological Reviews, 47, pp. 579-595Crooks, G.E., Hon, G., Chandonia, J.M., Brenner, S.E., WebLogo: a sequence logo generator (2004) Genome Research, 14, pp. 1188-1190Edgar, R.C., MUSCLE: multiple sequence alignment with high accuracy and high throughput (2004) Nucleic Acids Research, 32, pp. 1792-1797Fassbinder, F., Kist, M., Bereswill, S., Structural and functional analysis of the riboflavin synthesis genes encoding GTP cyclohydrolase II (ribA), DHBP synthase (ribBA), riboflavin synthase (ribC), and riboflavin deaminase/reductase (ribD) from Helicobacter pylori strain P1 (2000) FEMS Microbiology Letters, 191, pp. 191-197Felsenstein, J., PHYLIP - phylogeny inference package (version 3.2) (1989) Cladistics, 5, pp. 164-166Fox, B., Walsh, C.T., Mercuric reductase. Purification and characterization of a transposon-encoded flavoprotein containing an oxidation-reduction-active disulfide (1982) Journal of Biological Chemistry, 257, pp. 2498-2503Garcia Jr., O., Silva, L.L., Differences in growth and iron oxidation among Thiobacillus ferrooxidans cultures in the presence of some toxic metals (1991) Biotechnology Letters, 13, pp. 567-570Gelfand, M.S., Mironov, A.A., Jomantas, J., Kozlov, Y.I., Perumov, D.A., A conserved RNA structure element involved in the regulation of bacterial riboflavin synthesis genes (1999) Trends in Genetics, 15, pp. 439-442Griffiths-Jones, S., Moxon, S., Marshall, M., Khanna, A., Eddy, S.R., Bateman, A., Rfam: annotating non-coding RNAs in complete genomes (2005) Nucleic Acids Research, 33, pp. D121-D124Iwahori, K., Takeuchi, F., Kamimura, K., Sugio, T., Ferrous iron-dependent volatilization of mercury by the plasma membrane of Thiobacillus ferrooxidans (2000) Applied and Enviromental Microbiology, 66, pp. 3823-3827Kasai, S., Sumimoto, T., Stimulated biosynthesis of flavins in Photobacterium phosphoreum IFO 13896 and the presence of complete rib operons in two species of luminous bacteria (2002) European Journal of Biochemistry, 269, pp. 5851-5860Kelly, D.P., Wood, A.P., Reclassification of some species of Thiobacillus to the newly designated genera Acidithiobacillus gen. nov., Halothiobacillus gen. nov. and Thermithiobacillus gen. nov (2000) International Journal of Systematic and Evolutionary Microbiology, 50, pp. 511-516Kuenen, J.G., Tuovinen, O.H., The genera Thiobacillus and Thiomicrospira (1981) The prokaryotes, pp. 1023-1036. , Starr M.P., Stolp H., Truper H.G., Balows A., and Schegel H.G. (Eds), Springer-Verlag, BerlinLiao, D.I., Calabrese, J.C., Wawrzak, Z., Viitanen, P.V., Jordan, D.B., Crystal structure of 3,4-dihydroxy-2-butanone 4-phosphate synthase of riboflavin biosynthesis (2001) Structure, 9 (1), pp. 11-18Paulino, L.C., Mello, M.P., Ottoboni, L.M.M., Differential gene expression in response to copper in Acidithiobacillus ferrooxidans analyzed by RNA arbitrarily primed polymerase chain reaction (2002) Electrophoresis, 23, pp. 520-527Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular cloning: a laboratory manual, , Cold Spring Harbor Laboratory, Cold Spring Harbor, NYShiratori, T., Inoue, C., Sugawara, K., Kusano, T., Kitagawa, Y., Cloning and expression of Thiobacillus ferrooxidans mercury ion resistance genes in Escherichia coli (1989) Journal of Bacteriology, 171, pp. 3458-3464Siddharthan, R., Siggia, E.D., van Nimwegen, E., PhyloGibbs: a gibbs sampling motif finder that incorporates phylogeny (2005) PLoS Computational Biology 1, , e67Swofford, D.L., (2002) PAUP*: phylogenetic analysis using parsimony (*and other methods) version 4.0b 10, , Sinauer Associates, Sunderland, MAThompson, J.D., Higgins, D.G., Gibson, T.J., CLUSTAL W: improving the sensitivity of progressive multiple sequence alignments through sequence weighting, position specific gap penalties and weight matrix choice (1994) Nucleic Acids Research, 22, pp. 4673-4680Tuovinen, O.H., Kelly, D.P., Biology of Thiobacillus ferrooxidans in relation to the microbiological leaching of sulphide ore (1972) Zeitschrift für Allgemeine Mikrobiologie, 12, pp. 311-346Valdés, J., Veloso, F., Jedlicki, E., Holmes, D., Metabolic reconstruction of sulfur assimilation in the extremophile Acidithiobacillus ferrooxidans based on genome analysis (2003) BMC Genomics, 4, pp. 51-67Vitreschak, A.G., Rodionov, D.A., Mironov, A.A., Gelfand, M.S., Regulation of riboflavin biosynthesis and transport genes in bacteria by transcriptional and translational attenuation (2002) Nucleic Acids Research, 30, pp. 3141-3151Winderickx, J., Castro, J.M. (1994) Practical course in molecular biology of microorganisms, January 23-February 11, Universidade Federal de Ouro Preto - MG, pp. 59Winkler, W.C., Cohen-Chalamish, S., Breaker, R.R., An mRNA structure that controls gene expression by binding FMN (2002) Proceedings of the National Academy of Sciences USA, 99, pp. 15908-15913Worst, D.J., Gerrits, M.M., Vandenbroucke-Grauls, C.M., Kusters, J.G., Helicobacter pylori ribBA-mediated riboflavin production is involved in iron acquisition (1998) Journal of Bacteriology, 180, pp. 1473-1479Yarzábal, A., Brasseur, G., Bonnefoy, V., Cytochromes c of Acidithiobacillus ferrooxidans (2002) FEMS Microbiology Letters, 209, pp. 189-195Yarzábal, A., Brasseur, G., Ratouchniak, J., Lund, K., Lemesle-Meunier, D., DeMoss, J.A., The high molecular weight cytochrome c Cyc2 of Acidithiobacillus ferrooxidans is an outer membrane protein (2002) Journal of Bacteriology, 184, pp. 313-317Zuker, M., Mathews, D.H., Turner, D.H., Algorithms and thermodynamics for RNA secondary structure prediction: a practical guide (1999) RNA biochemistry and biotechnology, number 70 in Nato Science Series, pp. 11-43. , Barciszewski J., and Clark B.F.C. (Eds), Kluwer Academic Publishers, Dordrecht, The Netherland

Structural Aspects Of The P.p222q Homozygous Mutation Of Hsd3b2 Gene In A Patient With Congenital Adrenal Hyperplasia [aspectos Estruturais Da Mutação Homozigótica P.p222q Do Gene Hsd3b2 Em Um Paciente Com Hiperplasia Congênita Da Adrenal]

Type II 3β-hydroxysteroid dehydrogenase/D5-D4-isomerase (3β-HSD2), encoded by the HSD3B2 gene, is a key enzyme involved in the biosynthesis of all the classes of steroid hormones. Deleterious mutations in the HSD3B2 gene cause the classical deficiency of 3β-HSD2, which is a rare autosomal recessive disease that leads to congenital adrenal hyperplasia (CAH). CAH is the most frequent cause of ambiguous genitalia and adrenal insufficiency in newborn infants with variable degrees of salt losing. Here we report the molecular and structural analysis of the HSD3B2 gene in a 46, XY child, who was born from consanguineous parents, and presented with ambiguous genitalia and salt losing. The patient carries a homozygous nucleotide c.665C>A change in exon 4 that putatively substitutes the proline at codon 222 for glutamine. Molecular homology modeling of normal and mutant 3β-HSD2 enzymes emphasizes codon 222 as an important residue for the folding pattern of the enzyme and validates a suitable model for analysis of new mutations. © ABE&M todos os direitos reservados.548768774Simard, J., Durocher, F., Mebarki, F., Turgeon, C., Sanchez, R., Labrie, Y., Molecular biology and genetics of the 3b-hydroxysteroid dehydrogenase/D5-D4 isomerase gene family (1996) J Endocrin, 150, pp. S189-S207Labrie, F., Simard, J., Luu-The, V., Belanger, A., Pelletier, G., Structure, function and tissue-specific gene expression of 3b-hydroxysteroid dehydrogenase/D5-D4 isomerase enzymes in classical and peripheral intracrine steroidogenic tissues (1992) J Steroid Biochem Molec Biol, 43, pp. 805-826Thomas, J., Duax, W., Addlagatta, A., Brandt, S., Fuller, R., Norris, W., Structure/function relationships responsible for coenzyme specificity and the isomerase activity of human type 1 3 beta-hydroxysteroid dehydrogenase/isomerase (2003) J Biol Chem, 12 (37), pp. 35483-35490McBride, M., McVie, A., Burridge, S., Brintnell, B., Craig, N., Wallace, A., Cloning, expression, and physical mapping of the 3β-hydroxysteroid dehydrogenase gene cluster (HSD3BP1-HSD3BP5) in human (1999) Genomics, 61, pp. 277-284Lachance, Y., Luu-The, V., Verreault, H., Dumont, M., Rheaume, E., Leblanc, G., Structure of the human type II 3β-hydroxysteroid dehydrogenase/Δ5- Δ4 isomerase (3β-HSD) gene: Adrenal and gonadal specifity (1991) DNA and Cell Biol, 10, pp. 701-711Rheaume, E., Lachance, Y., Zhao, H., Breton, N., Dumont, M., de Launoit, Y., Structure and expression of a new complementary DNA encoding the almost exclusive 3β-hydroxysteroid dehydrogenase/ Δ5-Δ4 isomerase in human adrenals and gonads (1991) Mol Endocrinol, 8, pp. 1147-1157Rainey, W., Parker, C., Rehman, K., Carr, B., The adrenal genetic puzzle: How do the fetal and adult pieces differ? (2002) Endocr Res, 28 (4), pp. 611-622Rheaume, E., Simard, J., Morel, Y., Mebarki, F., Zachmann, M., Forest, M., Congenital adrenal hyperplasia due to point mutations in the type II 3β-hydroxysteroid dehydrogenase gene (1992) Nature Gen, 1, pp. 239-245Simard, J., Rheaume, E., Sanchez, R., Laflamme, N., Launoit, Y., Luu--the, V., Molecular basis of congenital adrenal hyperplasia due to 37beta;-hydroxysteroid dehydrogenase deficiency (1993) Mol Endocrinol, 7, pp. 716-728Pang, S., Congenital adrenal hyperplasia owing to 3β-hydroxysteroid dehydrogenase deficiency (2001) Endocrinol Metab Clin North Am, 1, pp. 81-99Simard, J., Moisan, A., Morel, Y., Congenital adrenal hyperplasia due to 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase deficiency (2002) Semin Reprod Med, 3, pp. 255-276Pang, S., Lerner, A.J., Stoner, E., Levine, L.S., Oberfield, S.E., Engel, I., Late-onset adrenal steroid 3 beta-hydroxysteroid dehydrogenase deficiency. I. A cause of hirsutism in pubertal and postpubertal women (1985) J Clin Endocrinol Metab, 60, pp. 428-439Schram, P., Zerah, M., Mani, P., Jewelewicz, R., Jaffe, S., New, M.I., Nonclassical 3beta-hydroxysteroid dehydrogenase deficiency: A review of our experience with 25 female patients (1992) Fertil Steril, 58, pp. 129-136Medina, M., Herrera, J., Flores, M., Normal ovarian function in a mild form of late-onset 3 beta-hydroxysteroid dehydrogenase deficiency (1986) Fertil Steril, 46, pp. 1021-1025Eldar-Geva, T., Hurwitz, A., Vecsei, P., Palti, Z., Milwidsky, A., Rösler, A., Secondary biosynthetic defects in women with women with late-onset congenital adrenal hyperplasia (1990) N Engl J Med, 323, pp. 855-863Zerah, M., Rhéaume, E., Mani, P., Schram, P., Simard, J., Labrie, F., No evidence of mutations in the genes for type I and type II 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) in nonclassical 3 beta HSD deficiency (1994) J Clin Endocrinol Metab, 79, pp. 1811-1817Chang, Y.T., Zhang, L., Sakkal-Aladdour, H., Absence of molecular defect in type II 3b-hydroxysteroid dehydrogenase (3bHSD) gene in premature pubarche children and hirsute female patients with moderately decreased adrenal 3bHSD activity (1995) Pediatr Res, 37, pp. 820-824Sakkal-Alkaddour, H., Zhang, L., Yang, X., Studies of 3b-hyddroxysteroid dehydrogenase genes in infants and children manifesting premature pubarche and increased ACTH stimulation D5 steroid levels (1996) J Clin Endocrinol Metab, 81, pp. 3961-3965Thoden, J., Frey, P., Holden, H., Crystal structures of the oxidized and reduced forms of UDP-galactose 4-epimerase isolated from Escherichia coli (1996) Biochemistry, 35, pp. 2557-2566Neshich, G., Borro, L., Higa, R., Kuser, P., Yamagishi, M., Franco, E., The Diamond STING server (2005) Nucleic Acids Res, 33 (web server issue), pp. W29-W35. , http://sms.cbi.cnptia.embrapa.brMendonca, B., Russel, A., Vasconcelos-Leite, M., Arnhold, I., Bloise, W., Wajchenberg, B., Mutation in 3β-hydroxysteroid dehydrogenase type II associated with pseudohermaphroditism in males and premature pubarche or cryptic expression in females (1994) J Mol Endocrinol, 12, pp. 119-122Moisan, A., Ricketts, M., Tardy, V., Desrochers, M., Mebarki, F., Chaussain, J., New insight into the molecular basis of 3beta-hydroxysteroid dehydrogenase deficiency: Identification of eight mutations in the HSD3B2 gene eleven patients from seven new families and comparison of the functional properties of twenty-five mutant enzymes (1999) J Clin Endocrinol Metab, 12, pp. 4410-4425Pang, S., Wang, W., Rich, B., David, R., Chang, Y., Carbunaru, G., A novel nonstop mutation in the stop codon and a novel missense mutation in the type II 3b-hydroxysteroid dehydrogenase (3b-HSD) gene causing, respectively, nonclassic and classic 3beta-HSD deficiency congenital adrenal hyperplasia (2002) J Clin Endocrinol Metab, 6, pp. 2556-2563Marui, S., Castro, M., Latronico, A.C., Elias, L.L., Arnhold, I.J., Moreira, A.C., Mutations in the type II 3beta-hydroxysteroid dehydrogenase (HSD3B2) gene can cause premature pubarche in girls (2000) Clin Endocrinol, 52 (1), pp. 67-75Mermejo, L., Elias, L., Marui, S., Moreira, A., Mendonca, B., de Castro, M., Refining hormonal diagnosis of type II 3beta-hydroxysteroid dehydrogenase deficiency in patients with premature pubarche and hirsutism based on HSD3B2 genotyping (2005) J Clin Endocrinol Metab, 90 (3), pp. 1287-1293Thomas, J., Nash, W., Myers, R., Crankshaw, M., Strickler, R., Affinity radiolabeling identifies peptides and amino acids associated with substrate binding in human placental 3b-hydroxy-D5-steroid dehydrogenase (1993) J Biol Chem, 268, pp. 18507-18512Borman, S., Proteomics: Taking over where genomics leaves off (2000) Chem Eng News, 78, pp. 31-37Sali, A., Target practice (2001) Nat Struct Biol, 8, pp. 482-484Vitkup, D., Melamud, E., Moult, J., Sander, C., Completeness in structural genomics (2001) Nat Struct Biol, 8, pp. 559-566Thomas, J., Duax, W., Addlagatta, A., Kacsoh, B., Brandt, S., Norris, W., Structure/function aspects of human 3β-hydroxysteroid dehydrogenase (2004) Mol Cell Endocrinol, 215, pp. 73-8

Differentially Expressed Genes In Response To Amoxicillin In Helicobacter Pylori Analyzed By Rna Arbitrarily Primed Pcr

Because the molecular mechanism of amoxicillin resistance in Helicobacter pylori seems to be partially explained by several mutational changes in the pbp1A gene, the aim of the present study was to evaluate the gene expression pattern in response to amoxicillin in the AmxR Hardenberg strain using RNA arbitrarily primed PCR (RAP-PCR). In the experiments, c. 100 differentially expressed RAP-PCR products were identified using five arbitrary primers. The cDNAs that presented the highest levels of induction or repression were cloned and sequenced, and the sequences were compared with those present in databases using the blast search algorithm. The differential expression of the isolated cDNAs was confirmed by real-time PCR. The preliminary results showed that amoxicillin alters the expression of five cDNAs involved in biosynthesis, two involved with pathogenesis, four related to cell envelope formation, two involved in cellular processes, three related with transport and binding proteins, one involved with protein degradation, one involved with energy metabolism and seven hypothetical proteins. Further analysis of these cDNAs will allow a better comprehension of both the molecular mechanism(s) of amoxicillin resistance and the adaptative mechanism(s) used by H. pylori in the presence of this antibiotic. © 2006 Federation of European Microbiological Societies.502226230Boigegrain, R.A., Salhim, I., Alvarez-Martinezm, M.T., MacHold, J., Fedon, Y., Arpagaus, M., Weise, C., Rouot, B., Release of periplasmic proteins of Brucella suis upon acidic shock involves the outer membrane protein Omp25 (2004) Infect Immun, 72, pp. 5693-5703Chambers, H.F., Methicillin resistance in staphylococci: Molecular and biochemical basis and clinical implications (1997) Clin Microbiol Rev, 10, pp. 781-791Contreras, M., Thiberge, J.M., Mandrand-Berthelot, M.A., Labigne, A., Characterization of the roles of NikR, a nickel-responsive pleiotropic autoregulator of Helicobacter pylori (2003) Mol Microbiol, 49, pp. 947-963Dore, M.P., Graham, D.Y., Sepulveda, A.R., Different penicillin-binding protein profiles in amoxicillin-resistant Helicobacter pylori (1999) Helicobacter, 4, pp. 154-161Dunn, B.E., Cohen, H., Blaser, M.J., Helicobacter pylori (1997) Clin Microbiol Rev, 10, pp. 720-741Frere, J.M., Joris, B., Granier, B., Matagne, A., Jacob, F., Bourguignon-Bellefroid, C., Diversity of the mechanisms of resistance to beta-lactam antibiotics (1991) Res Microbiol, 142, pp. 705-710Frias-Lopez, J., Bonheyo, G.T., Fouke, B.W., Identification of differential gene expression in bacteria associated with coral black band disease by using RNA-arbitrarily primed PCR (2004) Appl Environ Microbiol, 70, pp. 3687-3694Gerrits, M.M., Schuijffel, D., Van Zwet, A.A., Kuipers, E.J., Vandenbroucke-Grauls, C.M., Kusters, J.G., Alterations in penicillin-binding protein 1A confer resistance to beta-lactam antibiotics in Helicobacter pylori (2002) Antimicrob Agents Chemother, 46, pp. 2229-2233Gerrits, M.M., Godoy, A.P., Kuipers, E.J., Ribeiro, M.L., Stoof, J., Mendonca, S., Van Vliet, A.H., Kusters, J.G., Multiple mutations in or adjacent to the conserved penicillin-binding protein motifs of the penicillin-binding protein 1A confer amoxicillin resistance to Helicobacter pylori (2006) Helicobacter, 11, pp. 181-187Glupczynski, Y., Megraud, F., Lopez-Brea, M., Andersen, L.P., European multicentre survey of in vitro antimicrobial resistance in Helicobacter pylori (2001) Eur J Clin Microbiol Infect Dis, 20, pp. 820-823Goh, E.B., Yim, G., Tsui, W., McClure, J., Surette, M.G., Davies, J., Transcriptional modulation of bacterial gene expression by subinhibitory concentrations of antibiotics (2002) Proc Natl Acad Sci USA, 99, pp. 17025-17030Kwon, D.H., Dore, M.P., Kim, J.J., Kato, M., Lee, M., Wu, J.Y., Al, E., High-level beta-lactam resistance associated with acquired multidrug resistance in Helicobacter pylori (2003) Antimicrob Agents Chemother, 47, pp. 2169-2178Li, S., Xiao, X., Li, J., Luo, J., Wang, F., Identification of genes regulated by changing salinity in the deep-sea bacterium hewanella sp. WP3 using RNA arbitrarily primed PCR (2006) Extremophiles, 10, pp. 97-104McGee, D.J., Mobley, H.L., Mechanisms of Helicobacter pylori infection: Bacterial factors (1999) Curr Top Microbiol Immunol, 241, pp. 155-180Mendonca, S., Ecclissato, C., Sartori, M.S., Godoy, A.P., Guerzoni, R.A., Degger, M., Pedrazzoli Jr., J., Prevalence of Helicobacter pylori resistance to metronidazole, clarithromycin, amoxicillin, tetracycline, and furazolidone in Brazil (2000) Helicobacter, 5, pp. 79-83Meyer, J.M., Silliman, N.P., Wang, W., Siepman, N.Y., Sugg, J.E., Morris, D., Al, E., Risk factors for Helicobacter pylori resistance in the United States: The surveillance of H. pylori antimicrobial resistance partnership (SHARP) study, 1993-1999 (2002) Ann Int Med, 136, pp. 13-24Mlynarczyk, G., Mlynarczyk, A., Jeljaszewicz, J., Epidemiological aspects of antibiotic resistance in respiratory pathogens (2001) Int J Antimicrob Agents, 18, pp. 497-502Okamoto, T., Yoshiyama, H., Nakazawa, T., Park, I.D., Chang, M.W., Yanai, H., Al, E., A change in PBP1 is involved in amoxicillin resistance of clinical isolates of Helicobacter pylori (2002) J Antimicrob Chemother, 50, pp. 849-856Paulino, L.C., De Mello, M.P., Ottoboni, L.M.M., Differential gene expression in response to copper in Acidithiobacillus ferrooxidans analyzed by RNA arbitrarily primed polymerase chain reaction (2002) Electrophoresis, 23, pp. 520-527Penston, J.G., McColl, K.E.L., Eradication of Helicobacter pylori: An objective assessment of current therapies (1997) Br J Clin Pharmacol, 43, pp. 223-243Pramanik, A., Braun, V., Albomycin uptake via a ferric hydroxamate transport system of Streptococcus pneumoniae R6 (2006) J Bacteriol, 188, pp. 3878-3886Siroy, A., Molle, V., Lemaitre-Guillier, C., Vallenet, D., Pestel-Caron, M., Cozzone, A.J., Jouenne, T., De, E., Channel formation by CarO, the carbapenem resistance-associated outer membrane protein of Acinetobacter baumannii (2005) Antimicrob Agents Chemother, 49, pp. 4876-4883Van Zwet, A.A., Vandenbroucke-Grauls, C.M., Thijs, J.C., Van Der Wouden, E.J., Gerrits, M.M., Kusters, J.G., Al, E., Stable amoxicillin resistance in Helicobacter pylori (1998) Lancet, 352, p. 1595Williams, J.G., Kubelik, A.R., Livak, K.J., Rafalski, J.A., Tingey, S.V., DNA polymorphisms amplified by arbitrary primers are useful as genetic markers (1990) Nucleic Acids Res, 18, pp. 6531-6535Winderickx, J., Castro, J.M., (1994), Pratical course in molecular biology of microrganisms. Universidade Federal de Ouro Preto - MG, January 23 - February 11, p. 59Zeth, K., Diederichs, K., Welte, W., Engelhardt, H., Crystal structure of Omp32, the anion-selective porin from Comamonas acidovorans, in complex with a periplasmic peptide at 2.1 a resolution (2000) Structure, 8, pp. 981-99

Ferric Iron Uptake Genes Are Differentially Expressed In The Presence Of Copper Sulfides In Acidithiobacillus Ferrooxidans Strain Lr

Acidithiobacillus ferrooxidans is one of the most widely used microorganisms in bioleaching operations to recover copper from low-grade copper sulfide ores. This work aimed to investigate the relative expression of genes related to the iron uptake system when A. ferrooxidans LR was maintained in contact with chalcopyrite or bornite as the sole energy source. Real-time quantitative PCR analysis revealed that the presence of bornite had no effect on the expression of seven genes related to the siderophore-mediated Fe(III) uptake system, while in the presence of chalcopyrite the expression of the genes was up-regulated. Bioinformatic analysis of the genomic region where these genes were found revealed the existence of three new putative DNA-binding sequences for the ferric iron uptake transcriptional regulator (Fur). Electrophoretic mobility shift assays demonstrated that a purified A. ferrooxidans His-tagged Fur protein was able to bind in vitro to each of these putative Fur boxes, suggesting that Fur regulated the expression of these genes. The expression of fur and two known Fur-regulated genes, mntH and dsrK, was also investigated in the presence of chalcopyrite. While the expression of fur and mntH was up-regulated, the expression of dsrK was down-regulated. The low amount of ferrous iron in the medium was probably responsible for the up-regulation of fur and the genes related to the siderophore-mediated Fe(III) uptake system when A. ferrooxidans LR was kept in the presence of chalcopyrite. A homology model of the A. ferrooxidans Fur was constructed and revealed that the putative DNA-binding surface presents conserved positively charged residues, supporting a previously suggested mode of interaction with DNA. The up-regulation of fur and the siderophore-mediated Fe(III) uptake genes, and the down-regulation of dsrK suggest that in the presence of chalcopyrite Fur acts as a transcription inducer and repressor. © 2010 Springer Science+Business Media B.V.993609617Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., Lipman, D.J., Gapped BLAST and PSI-BLAST: A new generation of protein database search programs (1997) Nucleic Acids Research, 25 (17), pp. 3389-3402. , DOI 10.1093/nar/25.17.3389Andrews, S.C., Robinson, A.K., Rodriguez-Quinones, F., Bacterial iron homeostasis (2003) FEMS Microbiology Reviews, 27 (2-3), pp. 215-237. , DOI 10.1016/S0168-6445(03)00055-XBairoch, A., Boeckmann, B., Ferro, S., Gasteiger, E., Swiss-Prot: Juggling between evolution and stability (2004) Brief Bioinform, 5, pp. 39-55. , 1:CAS:528:DC%2BD2cXjs1ahsbw%3D 10.1093/bib/5.1.39 15153305Bevilaqua, D., Diez-Perez, I., Fugivara, C.S., Sanz, F., Garcia Jr., O., Benedetti, A.V., Characterization of bornite (Cu5FeS4) electrodes in the presence of the bacterium Acidithiobacillus ferrooxidans (2003) Journal of the Brazilian Chemical Society, 14 (4), pp. 637-644Carlos, C., Reis, F.C., Vicentini, R., Madureira, D.J., Ottoboni, L.M.M., The rus operon genes are differentially regulated when Acidithiobacillus ferrooxidans LR is kept in contact with metal sulfides (2008) Curr Microbiol, 57, pp. 375-380. , 1:CAS:528:DC%2BD1cXhtVWktbrJ 10.1007/s00284-008-9208-7 18665419Delano, W.L., (2002) The PyMOL Molecular Graphics System on the World Wide Web, , http://www.pymol.orgEdgar, R.C., MUSCLE: Multiple sequence alignment with high accuracy and high throughput (2004) Nucleic Acids Research, 32 (5), pp. 1792-1797. , DOI 10.1093/nar/gkh340Ferraz, L.F.C., Verde, L.C.L., Reis, F.C., Alexandrino, F., Felício, A.P., Novo, M.T.M., Garcia Jr., O., Ottoboni, L.M.M., Gene expression modulation by chalcopyrite and bornite in Acidithiobacillus ferrooxidans (2010) Arch Microbiol, 192, pp. 531-540. , 1:CAS:528:DC%2BC3cXnsFGksb0%3D 10.1007/s00203-010-0584-6 20480358Garcia Jr., O., Isolation and purification of Thiobacillus ferrooxidans and Thiobacillus thiooxidans from some coal and uranium mines of Brazil (1991) Rev Microbiol, 22, pp. 1-6Gouet, P., Courcelle, E., Stuart, D.I., Metoz, F., ESPript: Analysis of multiple sequence alignments in PostScript (1999) Bioinformatics, 15 (4), pp. 305-308. , DOI 10.1093/bioinformatics/15.4.305Griggs, D.W., Tharp, B.B., Konisky, J., Cloning and promoter identification of the iron-regulated cir gene of Escherichia coli (1987) Journal of Bacteriology, 169 (12), pp. 5343-5352Hall, H.K., Foster, J.W., The role of Fur in the acid tolerance response of Salmonella typhimurium is physiologically and genetically separable from its role in iron acquisition (1996) Journal of Bacteriology, 178 (19), pp. 5683-5691Kehres, D.G., Zaharik, M.L., Finlay, B.B., Maguire, M.E., The NRAMP proteins of Salmonella typhimurium and Escherichia coli are selective manganese transporters involved in the response to reactive oxygen (2000) Molecular Microbiology, 36 (5), pp. 1085-1100. , DOI 10.1046/j.1365-2958.2000.01922.xLivak, K.J., Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method (2001) Methods, 25 (4), pp. 402-408. , DOI 10.1006/meth.2001.1262Makui, H., Roig, E., Cole, S.T., Helmann, J.D., Gros, P., Cellier, M.F.M., Identification of the Escherichia coli K-12 Nramp orthologue (MntH) as a selective divalent metal ion transporter (2000) Molecular Microbiology, 35 (5), pp. 1065-1078. , DOI 10.1046/j.1365-2958.2000.01774.xMarchler-Bauer, A., Anderson, J.B., Derbyshire, M.K., DeWeese-Scott, C., Gonzales, N.R., Gwadz, M., Hao, L., Bryant, S.H., CDD: A conserved domain database for interactive domain family analysis (2007) Nucleic Acids Research, 35 (SUPPL. 1), pp. D237-D240. , DOI 10.1093/nar/gkl951Paulino, L.C., De Mello, M.P., Ottoboni, L.M.M., Differential gene expression in response to copper in Acidithiobacillus ferrooxidans analyzed by RNA arbitrarily primed polymerase chain reaction (2002) Electrophoresis, 23 (4), pp. 520-527. , DOI 10.1002/1522-2683(200202)23:43.0.CO;2-RPires, R.H., Venceslau, S.S., Morais, F., Teixeira, M., Xavier, A.V., Pereira, I.A.C., Characterization of the Desulfovibrio desulfuricans ATCC 27774 DsrMKJOP complex - A membrane-bound redox complex involved in the sulfate respiratory pathway (2006) Biochemistry, 45 (1), pp. 249-262. , DOI 10.1021/bi0515265Pohl, E., Haller, J.C., Mijovilovich, A., Meyer-Klaucke, W., Garman, E., Vasil, M.L., Architecture of a protein central to iron homeostasis: Crystal structure and spectroscopic analysis of the ferric uptake regulator (2003) Molecular Microbiology, 47 (4), pp. 903-915. , DOI 10.1046/j.1365-2958.2003.03337.xQuatrini, R., Lefimil, C., Holmes, D.S., Jedlicki, E., The ferric iron uptake regulator (Fur) from the extreme acidophile Acidithiobacillus ferrooxidans (2005) Microbiology, 151 (6), pp. 2005-2015. , DOI 10.1099/mic.0.27581-0Quatrini, R., Jedlicki, E., Holmes, D.S., Genomic insights into the iron uptake mechanisms of the biomining microorganism Acidithiobacillus ferrooxidans (2005) Journal of Industrial Microbiology and Biotechnology, 32 (11-12), pp. 606-614. , DOI 10.1007/s10295-005-0233-2Quatrini, R., Lefimil, C., Veloso, F.A., Pedroso, I., Holmes, D.S., Jedlicki, E., Bioinformatic prediction and experimental verification of Fur-regulated genes in the extreme acidophile Acidithiobacillus ferrooxidans (2007) Nucleic Acids Research, 35 (7), pp. 2153-2166. , DOI 10.1093/nar/gkm068Rawlings, D.E., Heavy metal mining using microbes (2002) Annual Review of Microbiology, 56, pp. 65-91. , DOI 10.1146/annurev.micro.56.012302.161052Reents, H., Munch, R., Dammeyer, T., Jahn, D., Hartig, E., The Fnr regulon of Bacillus subtilis (2006) Journal of Bacteriology, 188 (3), pp. 1103-1112. , DOI 10.1128/JB.188.3.1103-1112.2006Rivas, M., Seeger, M., Holmes, D.S., Jedlicki, E., A Lux-like quorum sensing system in the extreme acidophile Acidithiobacillus ferrooxidans (2005) Biological Research, 38 (2-3), pp. 283-297Sali, A., Blundell, T.L., Comparative protein modelling by satisfaction of spatial restraints (1993) Journal of Molecular Biology, 234 (3), pp. 779-815. , DOI 10.1006/jmbi.1993.1626Schneider, T.D., Spouge, J., Information content of individual genetic sequences (1997) Journal of Theoretical Biology, 189 (4), pp. 427-441. , DOI 10.1006/jtbi.1997.0540Third, K.A., Cord-Ruwisch, R., Watling, H.R., The role of iron-oxidizing bacteria in stimulation or inhibition of chalcopyrite bioleaching (2000) Hydrometallurgy, 57, pp. 225-233. , 1:CAS:528:DC%2BD3cXlsVKrtL0%3D 10.1016/S0304-386X(00)00115-8Tuovinen, O.H., Kelly, D.P., Studies on the growth of Thiobacillus ferrooxidans. I. Use of membrane filters and ferrous iron agar to determine viable numbers, and comparison with 14 CO2-fixation and iron oxidation as measures of growth (1973) Arch Mikrobiol, 88, pp. 285-298. , 1:CAS:528:DyaE3sXmvFWltA%3D%3D 10.1007/BF00409941 4684598Vogel, A.I., (1981) Análise Inorgânica Quantitative, , 4 edn. Guanabara Kogan, Rio de JaneiroYarzabal, A., Appia-Ayme, C., Ratouchniak, J., Bonnefoy, V., Regulation of the expression of the Acidithiobacillus ferrooxidans rus operon encoding two cytochromes c, a cytochrome oxidase and rusticyanin (2004) Microbiology, 150 (7), pp. 2113-212

46,xx Dsd And Antley-bixler Syndrome Due To Novel Mutations In The Cytochrome P450 Oxidoreductase Gene [dds 46,xx E Síndrome De Antley-bixler Causada Por Novas Mutações No Gene Da Enzima P450 Oxidorredutase]

Deficiency of the enzyme P450 oxidoreductase is a rare form of congenital adrenal hyperplasia with characteristics of combined and partial impairments in steroidogenic enzyme activities, as P450 oxidoreductase transfers electrons to CYP21A2, CYP17A1, and CYP19A1. It results in disorders of sex development and skeletal malformations similar to Antley-Bixley syndrome. We report the case of a 9-year-old girl who was born with virilized genitalia (Prader stage V), absence of palpable gonads, 46,XX karyotype, and hypergonadotropic hypogonadism. During the first year of life, ovarian cyst, partial adrenal insufficiency, and osteoarticular changes, such as mild craniosynostosis, carpal and tarsal synostosis, and limited forearm pronosupination were observed. Her mother presented severe virilization during pregnancy. The molecular analysis of P450 oxidoreductase gene revealed compound heterozygosis for the nonsense p.Arg223*, and the novel missense p.Met408Lys, inherited from the father and the mother, respectively. © ABEM todos os direitos reservados.568578585Miller, W.L., Minireview: Regulation of steroidogenesis by electron transfer (2005) Endocrinology, 146, pp. 2544-2550Scott, R.R., Gomes, L.G., Huang, N., Van Vliet, G., Miller, W.L., Apparent manifesting heterozygosity in P450 oxidoreductase deficiency and its effect on coexisting 21-hydroxylase deficiency (2007) J Clin Endocrinol Metab, 92, pp. 2318-2322Auchus, R.J., Lee, T.C., Miller, W.L., Cytochrome b5 augments the 17,20-lyase activity of human P450c17 without direct electron transfer (1998) J Biol Chem, 273, pp. 3158-3165Flück, C.E., Tajima, T., Pandey, A.V., Arlt, W., Okuhara, K., Verge, C.F., Mutant P450 oxidoreductase causes disordered steroidogenesis with and without Antley-Bixler syndrome (2004) Nat Genet, 36, pp. 228-230Arlt, W., Walker, E.A., Draper, N., Ivison, H.E., Ride, J.P., Hammer, F., Congenital adrenal hyperplasia caused by mutant P450 oxidoreductase and human androgen synthesis: Analytical study (2004) Lancet, 363, pp. 2128-2135Scott, R.R., Miller, W.L., Genetic and clinical features of P450 oxidoreductase deficiency (2008) Horm Res, 69, pp. 266-275Polusani, S.R., Kar, R., Riquelme, M.A., Masters, B.S., Panda, S.P., Regulation of gap junction function and connexin 43 expression by cytochrome P450 oxidoreductase (CYPOR) (2011) Biochem Biophys Res Commun, 411, pp. 490-495Tomalik-Scharte, D., Maiter, D., Kirchheiner, J., Ivison, H.E., Fuhr, U., Arlt, W., Impaired hepatic drug and steroid metabolism in congenital adrenal hyperplasia due to P450 oxidoreductase deficiency (2010) Eur J Endocrinol, 163, pp. 919-924(2012) Human Gene Mutation Database, , http://www.hgmd.cf.ac.uk/ac/index.php, Available at, Accessed on: Nov 17Flück, C.E., Pandey, A.V., Clinical and biochemical consequences of P450 oxidoreductase deficiency (2011) Endocr Dev, 20, pp. 63-79Krone, N., Reisch, N., Idkowiak, J., Dhir, V., Ivison, H.E., Hughes, B.A., Genotype-phenotype analysis in congenital adrenal hyperplasia due to P450 oxidoreductase deficiency (2012) J Clin Endocrinol Metab, 97, pp. E257-E267Sambrook, J., Fristsch, E.F., Maniatis, T.E., (1989) Molecular Cloning: A laboratory manual, , Cold Spring Harbor, NY: Cold Spring Harbor Laboratory PressXia, C., Panda, S.P., Marohnic, C.C., Martásek, P., Masters, B.S., Kim, J.J., Structural basis for human NADPH-cytochrome P450 oxidoreductase deficiency (2011) Proc Natl Acad Sci U S A, 108, pp. 13486-13491Idkowiak, J., O'Riordan, S., Reisch, N., Malunowicz, E.M., Collins, F., Kerstens, M.N., Pubertal presentation in seven patients with congenital adrenal hyperplasia due to P450 oxidoreductase deficiency (2011) J Clin Endocrinol Metab, 96 (3), pp. E453-E462Hamdane, D., Xia, C., Im, S.C., Zhang, H., Kim, J.J., Waskell, L., Structure and function of an NADPH-cytochrome P450 oxidoreductase in an open conformation capable of reducing cytochrome P450 (2009) J Biol Chem, 284 (17), pp. 11374-11384Xia, C., Hamdane, D., Shen, A.L., Choi, V., Kasper, C.B., Pearl, N.M., Conformational changes of NADPH-cytochrome P450 oxidoreductase are essential for catalysis and cofactor binding (2011) J Biol Chem, 286, pp. 16246-16260Herkert, J.C., Blaauwwiekel, E.E., Hoek, A., Veenstra-Knol, H.E., Kema, I.P., Arlt, W., A rare cause of congenital adrenal hyperplasia: Antley-Bixler syndrome due to POR deficiency (2011) Neth J Med, 69, pp. 281-283Fukami, M., Hasegawa, T., Horikawa, R., Ohashi, T., Nishimura, G., Homma, K., Cytochrome P450 oxidoreductase deficiency in three patients initially regarded as having 21-hydroxylase deficiency and/or aromatase deficiency: Diagnostic value of urine steroid hormone analysis (2006) Pediatr Res, 59, pp. 276-280Flück, C.E., Miller, W.L., P450 oxidoreductase deficiency: A new form of congenital adrenal hyperplasia (2006) Curr Opin Pediatr, 18, pp. 435-441Homma, K., Hasegawa, T., Nagai, T., Adachi, M., Horikawa, R., Fujiwara, I., Urine steroid hormone profile analysis in cytochrome P450 oxidoreductase deficiency: Implication for the backdoor pathway to dihydrotestosterone (2006) J Clin Endocrinol Metab, 91, pp. 2643-2649But, W.M., Lo, I.F., Shek, C.C., Tse, W.Y., Lam, S.T., Ambiguous genitalia, impaired steroidogenesis, and Antley-Bixler syndrome in a patient with P450 oxidoreductase deficiency (2010) Hong Kong Med J, 16, pp. 59-62Iijima, S., Ohishi, A., Ohzeki, T., Cytochrome P450 oxidoreductase deficiency with Antley-Bixler syndrome: Steroidogenic capacities (2009) J Pediatr Endocrinol Metab, 22, pp. 469-475Ko, J.M., Cheon, C.K., Kim, G.H., Yoo, H.W., A case of Antley-Bixler syndrome caused by compound heterozygous mutations of the cytochrome P450 oxidoreductase gene (2009) Eur J Pediatr, 168, pp. 877-880Sahakitrungruang, T., Huang, N., Tee, M.K., Agrawal, V., Russell, W.E., Crock, P., Clinical, genetic, and enzymatic characterization of P450 oxidoreductase deficiency in four patients (2009) J Clin Endocrinol Metab, 94, pp. 4992-5000Adachi, M., Tachibana, K., Asakura, Y., Yamamoto, T., Hanaki, K., Oka, A., Compound heterozygous mutations of cytochrome P450 oxidoreductase gene (POR) in two patients with Antley-Bixler syndrome (2004) Am J Med Genet A, 128, pp. 333-339New, M.I., Nonclassical congenital adrenal hyperplasia and the polycystic ovarian syndrome (1993) Ann NY Acad Sci, 687, pp. 193-205Rosa, S., Duff, C., Meyer, M., Lang-Muritano, M., Balercia, G., Boscaro, M., P450c17 deficiency: Clinical and molecular characterization of six patients (2007) J Clin Endocrinol Metab, 92, pp. 1000-1007Shima, M., Tanae, A., Miki, K., Katsumata, N., Matsumoto, S., Nakajima, S., Mechanism for the development of ovarian cysts in patients with congenital lipoid adrenal hyperplasia (2000) Eur J Endocrinol, 142, pp. 274-279Belgorosky, A., Pepe, C., Marino, R., Guercio, G., Saraco, N., Vaiani, E., Hypothalamic-pituitary-ovarian axis during infancy, early and late prepuberty in an aromatase-deficient girl who is a compound heterozygote for two new point mutations of the CYP19 gene (2003) J Clin Endocrinol Metab, 88, pp. 5127-5131Fukami, M., Horikawa, R., Nagai, T., Tanaka, T., Naiki, Y., Sato, N., Cytochrome P450 oxidoreductase gene mutations and Antley-Bixler syndrome with abnormal genitalia and/or impaired steroidogenesis: Molecular and clinical studies in 10 patients (2005) J Clin Endocrinol Metab, 90, pp. 414-426Grondahl, C., Hansen, T.H., Marky-Nielsen, K., Ottesen, J.L., Hyttel, P., Human oocyte maturation in vitro is stimulated by meiosis-activating sterol (2000) Hum Reprod, 15 (SUPPL. 5), pp. 3-1