Cited by 242Iron Metabolism and DisordersNeurological diseases and metabolismFerroptosis and cancer prognosis

Overexpression of Wild Type and Mutated Human Ferritin H-chain in HeLa Cells

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Cozzi, Anna, Corsi, Barbara, Levi, Sonia, Santambrogio, Paolo, Albertini, Alberto, Arosio, Paolo·2000·Journal of Biological Chemistry

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 approximately 5-fold increase of IRPs activity, approximately 2.5-fold increase of transferrin receptor, approximately 1.8-fold increase in iron-transferrin iron uptake, and approximately 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 H(2)O(2) 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 the 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.

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  1. Hentze, M. W., and Kuhn, L. C. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 8175-8182
  2. Gossen, M., and Bujard, H. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 5547-5551

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Authors
Cozzi, A., Corsi, B., Levi, S., Santambrogio, P., Albertini, A., & Arosio, P.
Affiliations
  • IRCCS Ospedale San Raffaele (IT)
  • University of Brescia (IT)
Corresponding author
Arosio, Paolo
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2000-08
Volume
275
Issue
33
Pages
25122-25129
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2030928461
Cited by
242
References
36
Citations by year
2025: 4 · 2024: 5 · 2023: 8 · 2022: 4 · 2021: 8 · 2020: 9 · 2019: 7 · 2018: 3 · 2017: 8 · 2016: 7 · 2015: 8 · 2014: 14 · 2013: 9 · 2012: 10
FWCI
6.27
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • INFM-OAR (INFN Catania) (submittedVersion, cc-by)
Topics
  • Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
  • Life Sciences › Neuroscience › Neurology › Neurological diseases and metabolism
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Ferroptosis and cancer prognosis
Keywords
ferroxidase activity, iron metabolism, iron regulatory proteins, transferrin receptor, iron absorption, labile iron pool, oxidative stress resistance, cell growth, HeLa cells, tetracycline-responsive promoter
MeSH terms
Anti-Bacterial Agents, Apoferritins, Cell Division, Ceruloplasmin, Doxycycline, Enzyme-Linked Immunosorbent Assay, Ferritins, HeLa Cells, Humans, Hydrogen Peroxide, Iron, Mutation, Plasmids, Precipitin Tests, Promoter Regions, Genetic, Tetracycline, Time Factors, Transfection, Immunoblotting, Mutagenesis, DNA, Complementary, Oxidative Stress

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