Artículo

Este artículo es de Acceso Abierto y puede ser descargado en su versión final desde nuestro repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is the most frequent inborn error of metabolism, and accounts for 90-95% of CAH cases. The affected enzyme, P450C21, is encoded by the CYP21A2 gene, located together with a 98% nucleotide sequence identity CYP21A1P pseudogene, on chromosome 6p21.3. Even though most patients carry CYP21A1P-derived mutations, an increasing number of novel and rare mutations in disease causing alleles were found in the last years. In the present work, we describe five CYP21A2 novel mutations, p.R132C, p.149C, p.M283V, p.E431K and a frameshift g.2511_2512delGG, in four non-classical and one salt wasting patients from Argentina. All novel point mutations are located in CYP21 protein residues that are conserved throughout mammalian species, and none of them were found in control individuals. The putative pathogenic mechanisms of the novel variants were analyzed in silico. A three-dimensional CYP21 structure was generated by homology modeling and the protein design algorithm FoldX was used to calculate changes in stability of CYP21A2 protein. Our analysis revealed changes in protein stability or in the surface charge of the mutant enzymes, which could be related to the clinical manifestation found in patients. © 2011 Minutolo et al.

Registro:

Documento: Artículo
Título:Structure-based analysis of five novel disease-causing mutations in 21-hydroxylase-deficient patients
Autor:Minutolo, C.; Nadra, A.D.; Fernández, C.; Taboas, M.; Buzzalino, N.; Casali, B.; Belli, S.; Charreau, E.H.; Alba, L.; Dain, L.
Filiación:Centro Nacional de Genética Médica, ANLIS, Buenos Aires, Argentina
Departamento de Fisiología Biología Molecular y Celular, Facultad de Ciencias Exactas y, Naturales Universidad de Buenos Aires, Buenos Aires, Argentina
Departamento de Química Bioló gica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Biología y Medicina Experimental, CONICET, Buenos Aires, Argentina
División Endocrinología, Hospital Durand, Buenos Aires, Argentina
Laboratorio de Citogené tica, Sión de Endocrinología, Hospital de Niños 'Dr. Ricardo Gutiérrez', Buenos Aires, Argentina
Palabras clave:cytochrome P450 2C; cytochrome P450 2C21; steroid 21 monooxygenase; unclassified drug; CYP21A2 protein, human; steroid 21 monooxygenase; article; chromosome 6p; congenital adrenal hyperplasia; exon; frameshift mutation; gene deletion; gene mutation; genotype; heterozygote detection; human; nucleotide sequence; phenotype; point mutation; promoter region; protein stability; protein structure; pseudogene; salt wasting; sequence analysis; steroid 21 monooxygenase deficiency; structure analysis; algorithm; Argentina; case control study; chemical structure; chemistry; congenital adrenal hyperplasia; genetic predisposition; genetics; mutation; Mammalia; Adrenal Hyperplasia, Congenital; Algorithms; Argentina; Case-Control Studies; Genetic Predisposition to Disease; Humans; Models, Molecular; Mutation; Protein Stability; Steroid 21-Hydroxylase
Año:2011
Volumen:6
Número:1
DOI: http://dx.doi.org/10.1371/journal.pone.0015899
Título revista:PLoS ONE
Título revista abreviado:PLoS ONE
ISSN:19326203
CAS:steroid 21 monooxygenase, 9029-68-9; CYP21A2 protein, human, 1.14.99.10; Steroid 21-Hydroxylase, 1.14.99.10
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_19326203_v6_n1_p_Minutolo.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v6_n1_p_Minutolo

Referencias:

  • New, M.I., White, P.C., Pang, S., Dupont, B., Speiser, P.W., The adrenal hyperplasias (1989) The Metabolic Basis of Inherited Disease, McGraw-Hill, pp. 1881-1917. , In: Scriver CR, Beaudet AL, Sly S, Valle D, eds., New York. 6th edn
  • Miller, W.L., Genetics, diagnosis and management of 21-hydroxylase deficiency (1994) J Clin Endocrinol Metab, 78, pp. 241-246
  • Pang, S., Shook, M.K., Current status of neonatal screening for congenital adrenal hyperplasia (1997) Curr Opin Pediatr, 9, pp. 419-423
  • Therell, B.L., Newborn screening for congenital adrenal hyperplasia (2001) Endocrinol Metab Clin North Am, 30, pp. 15-30
  • van der Kamp, H.J., Wit, J.M., Neonatal screening for congenital adrenal hyperplasia (2004) Eur J Endocrinol, 151, pp. U71-U75
  • Merke, D.P., Bornstein, S.R., Congenital adrenal hyperplasia (2005) Lancet, 365, pp. 2125-2136
  • Speiser, P.W., Dupont, B., Rubinstein, P., Piazza, A., Kastelan, A., New, M.I., High frequency of nonclassical steroid 21-hydroxylase deficiency (1985) Am J Hum Genet, 37, pp. 650-667
  • Donohoue, P.A., van Dop, C., McLean, R.H., Bias, W., Migeon, C.J., Gene conversion in salt-losing congenital adrenal hyperplasia with absent complement C4B protein (1986) J Clin Endocrinol Metab, 62, pp. 995-1002
  • Higashi, Y., Tanae, A., Inohue, H., Fujii-Kuriyama, Y., Evidence for frequent gene conversions in the steroid 21-hydroxylase (P-450c21) gene: Implications for 21-hydroxylase deficiency (1988) Am J Hum Genet, 42, pp. 17-25
  • Dain, L.B., Buzzalino, N.D., Oneto, A., Belli, S., Stivel, M., Classical and nonclassical 21-hydroxylase deficiency: A molecular study of Argentine patients (2002) Clin Endocrinol, 56, pp. 239-245
  • Pasqualini, T., Alonso, G., Tomasini, R., Galich, A.M., Buzzalino, N., Congenital adrenal hyperplasia: Clinical characteristics and genotype in newborn, childhood and adolescense (2007) Medicina, 67, pp. 253-261
  • den Dunnen, J.T., Antonarakis, S.E., Mutation nomenclature extensions and suggestions to describe complex mutations: A discussion (2000) Hum Mutat, 15, pp. 7-12
  • White, P.C., New, M.I., Dupont, B., Structure of the human steroid 21- hydroxylase genes (1986) Proc Natl Acad Sci USA, 83, pp. 5111-5115
  • Owerbach, D., Ballard, L., Draznin, M.B., Salt-wasting congenital adrenal hyperplasia: Detection and characterization of mutations in the steroid 21- hydroxylase gene, CYP21, using the polymerase chain reaction (1992) J Clin Endocrinol Metab, 74, pp. 553-558
  • Wedell, A., Luthman, H., Steroid 21-hydroxilase deficiency: Two additional mutations in salt-wasting disease and rapid screening of disease-causing mutations (1993) Hum Mol Genet, 2, pp. 499-504
  • Blanché, H., Vexiau, P., Clauin, S., le Gall, I., Fiet, J., Exhaustive screening of 21-hydroxylase gene in a population of hyperandrogenic women (1997) Hum Genet, 101, pp. 56-60
  • Dain, L., Minutolo, C., Buzzalino, N., Belli, S., Oneto, A., A novel CYP21A2 point mutation in a 21-hydroxylase deficient patient (2006) Novel Human Pathological Mutations. Hum. Genet, 119, pp. 359-364
  • Grünberg, R., Nilges, M., Leckner, J., Biskit - A software platform for structural bioinformatics (2007) Bioinformatics, 23, pp. 769-770
  • Schymkowitz, J., Borg, J., Stricher, F., Nys, R., Rousseau, F., Serrano, L., The FoldX web server: An online force field (2005) Nucleic Acids Res, 33, pp. W382-W388. , Web Server issue
  • Robins, T., Carlsson, J., Sunnerhagen, M., Wedell, A., Persson, B., Molecular Model of Human CYP21 Based on Mammalian CYP2C5: Structural Features Correlate with Clinical Severity of Mutations Causing Congenital Adrenal Hyperplasia (2006) Mol Endocrinol, 20, pp. 2946-2964
  • White, P.C., Speiser, P.W., Congenital adrenal hyperplasia due to 21- hydroxylase deficiency (2000) Endocrine Reviews, 21, pp. 245-291
  • Pey, A.L., Stricher, F., Serrano, L., Martinez, A., Predicted effects of missense mutations on native-state stability account for phenotypic outcome in phenylketonuria, a paradigm of misfolding diseases (2007) Am J Hum Genet, 81, pp. 1006-1024
  • Tusie-Luna, M.T., Traktman, P., White, P.C., Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus (1990) J Biol Chem, 265, pp. 20916-20922
  • Wu, D.A., Chung, B.C., Mutations of P450c21 (steroid 21-hydroxylase) at Cys428, Val281, and Ser268 result in complete, partial, or no loss of enzymatic activity, respectively (1991) J Clin Invest, 88, pp. 519-523
  • Watanabe, N., Kitazume, M., Fujisawaz, J., Mitsuaki, Y., Fujii-Kuriyama, Y.L., A novel CAMP-dependent regulatory region including a sequence like the CAMPresponsive element, far upstream of the human CYP21A2 gene (1993) Eur. J Biochem, 214, pp. 521-531
  • Wijesuriya, S.D., Zhang, G., Dardis, A., Miller, W.L., Transcriptional Regulatory Elements of the Human Gene for Cytochrome P450c21 (Steroid 21-Hydroxylase) Lie within Intron 35 of the Linked C4B Gene (1999) J Biol Chem, 274, pp. 38097-38106
  • Wilson, R.S., Mercado, A.B., Cheng, K.C., New, M.I., Steroid 21-hydroylase deficiency: Genotype may not predict phenotype (1995) J Clin Endocrinol Metab, 80, pp. 2322-2329
  • Jaaskelainen, J., Levo, A., Voutilainen, R., Partanen, J., Population-wide evaluation of disease manifestation in relation to molecular genotype in steroid 21-hydroxylase (CYP21) deficiency: Good correlation in a well defined population (1997) J Clin Endocrinol Metab, 82, pp. 3293-3297
  • Nimkarn, S., Cerame, B.I., Wei, J.Q., Dumic, M., Zunec, R., Congenital adrenal hyperplasia (21-hydroxylase deficiency) without demonstrable genetic mutations (1999) J Clin Endocrinol Metab, 84, pp. 378-381
  • Krone, N., Braun, A., Roscher, A.A., Knorr, D., Schwartz, H.P., Predicting phenotype in steroid 21-hydroxylase deficiency? Comprehensive genotyping in 155 unrelated, well defined patients from southern Germany (2000) J Clin Endocrinol Metab, 85, pp. 1059-1065
  • Ezquieta, B., Cueva, E., Varela, J., Oliver, A., Fernández, J., Non-classical 21-hydroxylase deficiency in children: Association of adrenocorticotropic hormone-stimulated 17-hydroxyprogesterone with the risk of compound heterozygosity with severe mutations (2002) Acta Paediatr, 91, pp. 892-898
  • Dolzan, V., Sólyom, J., Fekete, G., Kovács, J., Rakosnikova, V., Mutational spectrum of steroid 21-hydroxylase and the genotype-phenotype association in Middle European patients with congenital adrenal hyperplasia (2005) Eur J Endocrinol, 153, pp. 99-106
  • Gomes, L.G., Huang, N., Agrawal, V., Mendonça, B.B., Bachega, T.A.S.S., The Common P450 Oxidoreductase Variant A503V is Not a Modifier Gene for 21- Hydroxylase Deficiency (2008) J Clin Endocrinol Metab, 93, pp. 2913-2916
  • Hlavica, P., Schulze, J., Lewis, D.F., Functional interaction of cytochrome P450 with its redox partners: A critical assessment and update of the topology of predicted contact regions (2003) J Inorg Biochem, 96, pp. 279-297
  • Loidi, L., Quinteiro, C., Parajes, S., Barreiro, J., Leston, D.G., High variability in CYP21A2 mutated alleles in Spanish 21-hydroxylase deficiency patients, six novel mutations and a founder effect (2006) Clin Endocrinol, 64, pp. 330-336
  • Bleicken, C., Loidi, L., Dhir, V., Parajes, S., Quinteiro, C., Functional characterization of three CYP21A2 sequence variants (p.A265V, p.W302S, p.D322G) employing a yeast co-expression system (2008) Hum Mutat, 30, pp. 443-450
  • Nikoshkov, A., Lajic, S., Holst, M., Wedell, A., Luthman, H., Synergistic effect of partially inactivating mutations in steroid 21-hydroxylase deficiency (1997) J Clin Endocrinol Metab, 82, pp. 194-199
  • Menassa, R., Tardy, F., Despert, C., Bouvattier-Morel, J.P., Brossier, M., P.H62L a rare mutation of the CYP21 gene identified in two form of 21- hydroxylase deficiency (2008) J Clin Endocrinol Metab, 93, pp. 1901-1908
  • Soardi, F.C., Barbaro, M., Lau, I.F., Lemos-Marini, S.H., Baptista, M.T., Inhibition of CYP21A2 enzyme activity caused by novel missense mutations identified in Brazilian and Scandinavian patients (2008) J Clin Endocrinol Metab, 93, pp. 2416-2420
  • Tardy, V., Menassa, R., Sulmont, V., Lienhardt-Roussie, A., Lecointre, C., Phenotype-genotype correlations of 13 rare CYP21A2 mutations detected in 46 patients affected with 21-hydroxylase deficiency and in one carrier (2010) J Clin Endocrinol Metab, 95, pp. 1288-1300
  • Wedell, A., Ritzen, E.M., Haglund-Stengler, B., Luthman, H., Steroid21- hydroxylase deficiency: Three additional mutated alleles and establishment of phenotype-genotype relationships of common mutations (1992) Proc Natl Acad Sci USA, 89, pp. 7232-7236

Citas:

---------- APA ----------
Minutolo, C., Nadra, A.D., Fernández, C., Taboas, M., Buzzalino, N., Casali, B., Belli, S.,..., Dain, L. (2011) . Structure-based analysis of five novel disease-causing mutations in 21-hydroxylase-deficient patients. PLoS ONE, 6(1).
http://dx.doi.org/10.1371/journal.pone.0015899
---------- CHICAGO ----------
Minutolo, C., Nadra, A.D., Fernández, C., Taboas, M., Buzzalino, N., Casali, B., et al. "Structure-based analysis of five novel disease-causing mutations in 21-hydroxylase-deficient patients" . PLoS ONE 6, no. 1 (2011).
http://dx.doi.org/10.1371/journal.pone.0015899
---------- MLA ----------
Minutolo, C., Nadra, A.D., Fernández, C., Taboas, M., Buzzalino, N., Casali, B., et al. "Structure-based analysis of five novel disease-causing mutations in 21-hydroxylase-deficient patients" . PLoS ONE, vol. 6, no. 1, 2011.
http://dx.doi.org/10.1371/journal.pone.0015899
---------- VANCOUVER ----------
Minutolo, C., Nadra, A.D., Fernández, C., Taboas, M., Buzzalino, N., Casali, B., et al. Structure-based analysis of five novel disease-causing mutations in 21-hydroxylase-deficient patients. PLoS ONE. 2011;6(1).
http://dx.doi.org/10.1371/journal.pone.0015899