| Texto completo | |
| Autor(es): Mostrar menos - |
Madeira, Joao L. O.
;
Jorge, Alexander A. L.
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Martin, Regina M.
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Montenegro, Luciana R.
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Franca, Marcela M.
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Costalonga, Everlayny F.
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Correa, Fernanda A.
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Otto, Aline P.
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Arnhold, Ivo J. P.
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Freitas, Helayne S.
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Machado, Ubiratan F.
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Mendonca, Berenice B.
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Carvalho, Luciani R.
Número total de Autores: 13
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| Tipo de documento: | Artigo Científico |
| Fonte: | EUROPEAN JOURNAL OF ENDOCRINOLOGY; v. 175, n. 2, p. K7-K15, AUG 2016. |
| Citações Web of Science: | 2 |
| Resumo | |
Context: Mutations in the GH1 promoter are a rare cause of isolated growth hormone deficiency (IGHD). Objective: To identify the molecular aetiology of a family with IGHD. Design: DNA sequencing, electromobility shift (EMSA) and luciferase reporter assays. Setting: University Hospital. Patients: Three siblings (2M) born to consanguineous parents presented with IGHD with normal pituitary on MRI. Methods: The GH1 proximal promoter, locus control region, five exons and four introns as well as GHRHR gene were sequenced in genomic DNA by Sanger method. DNA- protein interaction was evaluated by EMSA in nuclear extracts of GH3 pituitary cells. Dual-luciferase reporter assays were performed in cells transiently transfected with plasmids containing four different combinations of GH1 allelic variants (AV). Results: The patients harboured two homozygous variants (c.-185T>C and c.-223C>T) in the GH1 promoter within a highly conserved region and predicted binding sites for POU1F1/SP1 and SP1 respectively. The parents and brother were carriers and these variants were absent in 100 controls. EMSA demonstrated absent binding of GH3 nuclear extract to the c.-223C>T variant and normal binding of both POU1F1 protein and GH3 nuclear extract to the c.-185T>C variant. In contrast to GH1 promoter with AV only at c.-185, the GH1 promoter containing the AV only at c.-223 and at both positions drove significantly less expression of luciferase compared with the promoter containing either positions wild type in luciferase reporter assays. Conclusion: To our knowledge, c.-223C>T is the first homozygous point mutation in the GH1 promoter that leads to short stature due to IGHD. (AU) | |
| Processo FAPESP: | 13/03236-5 - Novas abordagens e metodologias na investigação genético-molecular dos distúrbios de crescimento e desenvolvimento puberal |
| Beneficiário: | Alexander Augusto de Lima Jorge |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |
| Processo FAPESP: | 10/05188-0 - Validação da Dosagem de 11-desoxicortisol por Radioimunoensaio para Diagnóstico dos Distúrbios Hiperandrogênicos |
| Beneficiário: | João Luiz de Oliveira Madeira |
| Modalidade de apoio: | Bolsas no Brasil - Iniciação Científica |