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Influence of the polymorphisms of the alpha-major regulatory element HS-40 on in vitro gene expression
Ribeiro, D. M; Zaccariotto, T. R; Santos, M. N. N; Costa, F. F; Sonati, M. F.
  • Ribeiro, D. M; Universidade Estadual de Campinas. Departamento de Patologia Clínica. Campinas. BR
  • Zaccariotto, T. R; Universidade Estadual de Campinas. Departamento de Patologia Clínica. Campinas. BR
  • Santos, M. N. N; Universidade Estadual de Campinas. Departamento de Patologia Clínica. Campinas. BR
  • Costa, F. F; Universidade Estadual de Campinas. Faculdade de Ciências Médicas. Hemocentro e Departamento de Clínica Médica. Campinas. BR
  • Sonati, M. F; Universidade Estadual de Campinas. Departamento de Patologia Clínica. Campinas. BR
Braz. j. med. biol. res ; 42(9): 783-786, Sept. 2009. graf, tab
Article in English | LILACS | ID: lil-524320
ABSTRACT
The α-MRE is the major regulatory element responsible for the expression of human α-like globin genes. It is genetically polymorphic, and six different haplotypes, named A to F, have been identified in some population groups from Europe, Africa and Asia and in native Indians from two Brazilian Indian tribes. Most of the mutations that constitute the α-MRE haplotypes are located in flanking sequences of binding sites for nuclear factors. To our knowledge, there are no experimental studies evaluating whether such variability may influence the α-MRE enhancer activity. We analyzed and compared the expression of luciferase of nine constructs containing different α-MRE elements as enhancers. Genomic DNA samples from controls with A (wild-type α-MRE) and B haplotypes were used to generate C-F haplotypes by site-directed mutagenesis. In addition, three other elements containing only the G→A polymorphism at positions +130, +199, and +209, separately, were also tested. The different α-MRE elements were amplified and cloned into a plasmid containing the luciferase reporter gene and the SV40 promoter and used to transiently transfect K562 cells. A noticeable reduction in luciferase expression was observed with all constructs compared with the A haplotype. The greatest reductions occurred with the F haplotype (+96, C→A) and the isolated polymorphism +209, both located near the SP1 protein-binding sites believed not to be active in vivo. These are the first analyses of α-MRE polymorphisms on gene expression and demonstrate that these single nucleotide polymorphisms, although outside the binding sites for nuclear factors, are able to influence in vitro gene expression.
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Full text: Available Index: LILACS (Americas) Main subject: Polymorphism, Genetic / Haplotypes / Globins / Gene Expression Regulation / Regulatory Elements, Transcriptional / Mutation Type of study: Prognostic study Limits: Humans Language: English Journal: Braz. j. med. biol. res Journal subject: Biology / Medicine Year: 2009 Type: Article / Project document Affiliation country: Brazil Institution/Affiliation country: Universidade Estadual de Campinas/BR

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Full text: Available Index: LILACS (Americas) Main subject: Polymorphism, Genetic / Haplotypes / Globins / Gene Expression Regulation / Regulatory Elements, Transcriptional / Mutation Type of study: Prognostic study Limits: Humans Language: English Journal: Braz. j. med. biol. res Journal subject: Biology / Medicine Year: 2009 Type: Article / Project document Affiliation country: Brazil Institution/Affiliation country: Universidade Estadual de Campinas/BR