Identification of two novel mutations in the 5′untranslated region of H-ferritin using denaturing high performance liquid chromatography scanning

Laura Cremonesi, Barbara Foglieni, Isabella Fermo, Anna Cozzi, Rita Paroni, Giuseppina Ruggeri, Silvana Belloli, Sonia Levi, Silvia Fargion, Maurizio Ferrari, Paolo Arosio

Research output: Contribution to journalArticlepeer-review

Abstract

Background and Objectives. Hereditary hyperferritinemia cataract syndrome is caused by mutations of the iron responsive elements (IREs) of L-ferritin mRNA. These alter the IRE structure and determine L-ferritin upregulation. IREs are located in 5′ untranslated regions (5′ UTR) of ferritin mRNAs. L-ferritin 5′ UTR has been extensively studied and up to 21 different mutations have been identified. Only one mutation has been reported for H-ferritin 5′ UTR; this mutation modified IRE structure and was apparently associated with high serum ferritin levels and iron overload. Design and Methods. To identify other mutations in H ferritin 5′ UTR we developed a fast DNA scanning method based on denaturing high performance liquid chromatography (HPLC). Five artificial DNA mutants were produced in order to validate the analytical conditions of the system for the identification of all mutations by single runs at 68°C. The system was used to screen 660 DNA samples from subjects with high serum ferritin levels. Results. Two abnormal patterns were identified carrying the mutations C20G and G34T. Structural data and the analysis of ferritin levels in red blood cells suggest that these mutations do not affect the functionality of the IRE. Interpretation and Conclusions. This large and first population analysis indicates that mutations in the H-ferritin 5′ UTR are rare and do not seem to contribute to hyperferritinemia or iron overload.

Original languageEnglish
Pages (from-to)1110-1116
Number of pages7
JournalHaematologica
Volume88
Issue number10
Publication statusPublished - Oct 2003

Keywords

  • Denaturing HPLC
  • DNA variations
  • Ferritin
  • Hyperferritinemia
  • Iron metabolism

ASJC Scopus subject areas

  • Hematology

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