Please use this identifier to cite or link to this item:
http://repositorio.unicamp.br/jspui/handle/REPOSIP/195636
Type: | Artigo de periódico |
Title: | Analysis Of The Mechanism Of Action Of The Brazilian Type (agamma-195 C --> G) Of Hereditary Persistence Of Fetal Hemoglobin. |
Author: | Takahashi, T Schreiber, R Krieger, J E Saad, S T O Costa, F F |
Abstract: | We report an in vitro expression study of the Agamma-globin gene promoter containing the Agamma-195 C --> G mutation that causes the Brazilian type of hereditary persistence of fetal hemoglobin (HPFH). To demonstrate that this mutation results in increased promoter strength, we evaluated the mutant promoter linked to the hypersensitive site-2 of the locus control region with the luciferase reporter gene system and examined protein interactions by eletrophoretic mobility shift assay. The transient expression was studied in three cell lines: K562, HEL and 293, and indicated increased promoter activity of the promoter containing the Brazilian mutation in all cell lines. The protein-DNA interaction showed that, in contrast to the Agamma-198 T --> C mutation which has increased affinity for the Sp1 protein and creates a motif that behaves like a novel CACCC box in the gamma promoter, the Brazilian HPFH mutation decreases the affinity at the Sp1 protein and does not act as a CACCC motif. These results suggest that this mutation may act to increase the Agamma-globin chain production. In addition, the mechanism by which this increased production occurs is different to that of the -198 mutation. Other proteins may be involved in the overexpression of the gamma-globin chain and/or may be dependent upon the DNA structure. |
Subject: | Binding Sites Brazil Cell Line Cell Line, Transformed Dna Electrophoretic Mobility Shift Assay Embryo, Mammalian Fetal Hemoglobin Gene Expression Genes, Reporter Globins Hemoglobinopathies Humans Kidney Leukemia, Erythroblastic, Acute Luciferases Point Mutation Polymerase Chain Reaction Promoter Regions, Genetic Sp1 Transcription Factor Transfection Tumor Cells, Cultured |
Citation: | European Journal Of Haematology. v. 71, n. 6, p. 418-24, 2003-Dec. |
Rights: | fechado |
Address: | http://www.ncbi.nlm.nih.gov/pubmed/14703691 |
Date Issue: | 2003 |
Appears in Collections: | Unicamp - Artigos e Outros Documentos |
Files in This Item:
File | Size | Format | |
---|---|---|---|
pmed_14703691.pdf | 156.14 kB | Adobe PDF | View/Open |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.