title: Structural and Functional Insights on an Uncharacterized Aγ-Globin-Gene Polymorphism Present in Four β0-Thalassemia Families with High Fetal Hemoglobin Levels.
abstract: BACKGROUND
  Several DNA polymorphisms have been associated with high production of fetal hemoglobin (HbF), although the molecular basis is not completely understood. In order to identify and characterize novel HbF-associated elements, we focused on five probands and their four families (from Egypt, Iraq and Iran) with thalassemia major (either β(0)-IVSII-1 or β(0)-IVSI-1) and unusual HbF elevation (>98 %), congenital or acquired after rejection of bone marrow transplantation, suggesting an anticipated favorable genetic background to high HbF expression.
  
  
  METHODS
  Patient recruitment, genomic DNA sequencing, western blotting, electrophoretic mobility shift assays, surface plasmon resonance (SPR) biospecific interaction analysis, bioinformatics analyses based on docking experiments.
  
  
  RESULTS
  A polymorphism of the Aγ-globin gene is here studied in four families with β(0)-thalassemia (β(0)-IVSII-1 and β(0)-IVSI-1) and expressing unusual high HbF levels, congenital or acquired after rejection of bone marrow transplantation. This (G→A) polymorphism is present at position +25 of the Aγ-globin genes, corresponding to a 5'-UTR region of the Aγ-globin mRNA and, when present, is physically linked in chromosomes 11 of all the familiar members studied to the XmnI polymorphism and to the β(0)-thalassemia mutations. The region corresponding to the +25(G→A) polymorphism of the Aγ-globin gene belongs to a sequence recognized by DNA-binding protein complexes, including LYAR (Ly-1 antibody reactive clone), a zinc-finger transcription factor previously proposed to be involved in down-regulation of the expression of γ-globin genes in erythroid cells.
  
  
  CONCLUSIONS
  We found a novel polymorphism of the Aγ-globin gene in four families with β(0)-thalassemia and high levels of HbF expression. Additionally, we report evidence suggesting that the Aγ-globin gene +25(G→A) polymorphism decreases the efficiency of the interaction between this sequence and specific DNA binding protein complexes.
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Bianchi
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Lucia Carmela
authors.initials: LC
authors.lastname: Cosenza
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Lampronti
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Finotti
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Breveglieri
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Zuccato
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Fabbri
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padua, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Marzaro
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padua, Italy.
authors.forename: Adriana
authors.initials: A
authors.lastname: Chilin
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Gioia
authors.initials: G
authors.lastname: De Angelis
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Borgatti
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Cristiano
authors.initials: C
authors.lastname: Gallucci
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Alfieri
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Ribersani
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Isgrò
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Marziali
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Javid
authors.initials: J
authors.lastname: Gaziev
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Aldo
authors.initials: A
authors.lastname: Morrone
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Sodani
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Guido
authors.initials: G
authors.lastname: Lucarelli
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy. gam@unife.it.|Biotechnology Center, Ferrara University, Ferrara, Italy. gam@unife.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Gambari
authors.affiliation: International Centre for Transplantation in Thalassemia and Sickle Cell Anaemia, Mediterranean Institute of Haematology, Policlinic of "Tor Vergata" University, Rome, Italy.
authors.forename: Katia
authors.initials: K
authors.lastname: Paciaroni
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D005260:Female; D005319:Fetal Hemoglobin / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020014:K562 Cells; D008297:Male; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide; D017086:beta-Thalassemia / Q000235:genetics* / Q000378:metabolism; D055545:gamma-Globins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D055545:gamma-Globins; D005319:Fetal Hemoglobin