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| Author(s): |
Fernanda Marconi Roversi
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas |
| Defense date: | 2011-10-31 |
| Examining board members: |
Fernando Ferreira Costa;
Aderson da Silva Araujo;
Licio Augusto Velloso;
Fabíola Traina;
José Orlando Bordin
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| Advisor: | Anderson Ferreira da Cunha; Fernando Ferreira Costa |
| Abstract | |
Hemoglobin switching occurs after birth resulting in a decrease in gamma globin gene expression. Although, some single point mutation in this gene are able to maintain its expression during adult life, leading to increased production of fetal hemoglobin (HbF) which characterize the non deletion Hereditary Persistence of Fetal Hemoglobin (ndHPFH). Among these, the Brazilian type of ndHPFH corresponds to the C?G substitution at the -195 position of the A gamma globin gene. However, the molecular mechanism responsible for this phenotype still unclear. In contrast to the British ndHPFH type (-198), where the mechanism responsible for the increase of HbF levels is mediated by the raising in the affinity for the SP1 transcription factor, the Brazilian ndHPFH mutation does not affect Sp1 binding. In order to elucidate this mechanism, we tried to identify others transcription factors involved in the reactivation of the A gamma globin gene in the Brazilian ndHPFH. Alterations in the binding of two transcription factors were identified by DNA-protein Array, EMSA and ChIP: NF-E1/YY1 and PAX-1. Our results suggest that the -195C?G in the A gamma globin promoter may abrogate NF-E1/YY1 binding and increase PAX-1 binding in this DNA region, probably resulting in the reactivation of this gene. We also investigate others genes that may be involved in hemoglobin switching and/or maintenance of elevated HbF levels in Brazilian ndHPHF using Suppression Subtractive Hybridization Library to identify transcripts that are differentially expressed in reticulocytes of ndHPFH Brazilian subjects. MIER1 expression was found to be decreased in Brazilian ndHPFH reticulocytes, compared to controls. The MIER1 gene is able to chromatin-remodeling leading to the formation of heterochromatin and, consequently, silencing genes. This MIER1 may be an important gene in gamma to beta globin gene switching, where it could help in the maintenance of a closed chromatin structure in the gamma globin gene in individuals with low HbF levels. Another gene identified was KLF1 which expression was decreased in Brazilian HPFH and favors the interaction between the A gamma globin gene and the LCR. These results provide the first in vitro evidence for the possibly molecular mechanism of reactivation of the A gamma globin gene in Brazilian nHPFH. (AU) | |