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Functional analysis of a novel KLF1 gene promoter variation associated with hereditary persistence of fetal hemoglobin

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Abstract

Hereditary persistence of fetal hemoglobin (HPFH) is a rare hereditary condition resulting in elevated levels of fetal hemoglobin (HbF) in adults. Typical HPFH is associated with promoter mutations or large deletions affecting the human fetal globin (HBG1 and HBG2) genes, while genetic defects in other genes involved in human erythropoiesis, e.g. KLF1, also result in atypical HPFH. Here, we report the first KLF1 gene promoter mutation (KLF1:g.-148G > A) that is associated with increased HbF level. This mutation was shown to result in drastically reduced CAT reporter gene expression in K562 cells, compared to the wild-type sequence (p = 0.009) and also in reduced KLF1 gene expression in vivo. Furthermore, consistent with in silico analysis, electrophoretic mobility shift analysis showed that the KLF1:g.-148G > A mutation resides in a Sp1 binding site and further that this mutation leads to the ablation of Sp1 binding in vitro. These data suggest that the KLF1:g-148G > A mutation could play a role in increasing HbF levels in adults and further underlines the role of KLF1 as one of the key transcription factors involved in human fetal globin gene switching.

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Acknowledgments

This work was partly supported by grants from the Research Promotion Foundation (Cyprus, ΠΔΕ046_02) and the European Commission [International Thalassemia Network (ITHANET) Coordination action 026539] to GPP and by grant no. III41004 from Ministry of Education and Science, Republic of Serbia to SP.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, Laboratory for Molecular Hematology, University of Belgrade, Vojvode Stepe 444a, 11010, Belgrade, Serbia

    Milena Radmilovic, Branka Zukic, Maja Stojiljkovic Petrovic, Biljana Stankovic, Nikola Kotur & Sonja Pavlovic

  2. Department of Pharmacy, School of Health Sciences, University of Patras, University Campus, Rion, GR-26504, Patras, Greece

    Marina Bartsakoulia, Marianthi Georgitsi & George P. Patrinos

  3. University Children’s Hospital, School of Medicine, University of Belgrade, Belgrade, Serbia

    Lidija Dokmanovic

Authors
  1. Milena Radmilovic
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  2. Branka Zukic
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  3. Maja Stojiljkovic Petrovic
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  4. Marina Bartsakoulia
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  5. Biljana Stankovic
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  6. Nikola Kotur
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  7. Lidija Dokmanovic
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  8. Marianthi Georgitsi
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  9. George P. Patrinos
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  10. Sonja Pavlovic
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Corresponding authors

Correspondence to George P. Patrinos or Sonja Pavlovic.

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Radmilovic, M., Zukic, B., Petrovic, M.S. et al. Functional analysis of a novel KLF1 gene promoter variation associated with hereditary persistence of fetal hemoglobin. Ann Hematol 92, 53–58 (2013). https://doi.org/10.1007/s00277-012-1625-9

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  • DOI: https://doi.org/10.1007/s00277-012-1625-9

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