Overexpression of wild type and mutated human ferritin H-chain in HeLa cells: In vivo role of ferritin ferroxidase activity

A. Cozzi, B. Corsi, S. Levi, P. Santambrogio, A. Albertini, P. Arosio

Research output: Contribution to journalArticle

Abstract

Transfectant HeLa cells were generated that expressed human ferritin H-chain wild type and an H-chain mutant with inactivated ferroxidase activity under the control of the tetracycline-responsive promoter (Tet-off). The clones accumulated exogenous ferritins up to levels 14-16-fold over background, half of which were as H-chain homopolymers. This had no evident effect in the mutant ferritin clone, whereas it induced an iron-deficient phenotype in the H-ferritin wild type clone, manifested by ~5-fold increase of IRPs activity, ~2.5-fold increase of transferrin receptor, ~1.8-fold increase in iron-transferrin iron uptake, and ~50% reduction of labile iron pool. Overexpression of the H-ferritin, but not of the mutant ferritin, strongly reduced cell growth and increased resistance to H2O2 toxicity, effects that were reverted by prolonged incubation in iron-supplemented medium. The results show that in HeLa cells H-ferritin regulates the metabolic iron pool with a mechanism dependent on thai functionality of the ferroxidase centers, and this affects, in opposite directions, cellular growth and resistance to oxidative damage. This, and the finding that also in vivo H-chain homopolymers are much less efficient than the H/L heteropolymers in taking up iron, indicate that functional activity of H-ferritin in HeLa cells is that predicted from the in vitro data.

Original languageEnglish
Pages (from-to)25122-25129
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number33
DOIs
Publication statusPublished - Aug 18 2000

ASJC Scopus subject areas

  • Biochemistry

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