Metallothionein protects against the cytotoxic and DNA-damaging effects of nitric oxide

Margaret A. Schwarz, John S. Lazo, Jack C. Yalowich, William P. Allen, Mark Whitmore, Hector A. Bergonia, Edith Tzeng, Timothy R. Billiar, Paul D. Robbins, Jack R. Lancaster, Bruce R. Pitt*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

235 Scopus citations


In inflammatory states, nitric oxide (·NO) may be synthesized from precursor L-arginine via inducible ·NO synthase (iNOS) in large amounts for prolonged periods of time. When ·NO acts as an effector molecule under these conditions, it may be toxic to cells by inhibition of iron-containing enzymes or initiation of DNA single-strand breaks. In contrast to molecular targets of ·NO, considerably less is known regarding mechanisms by which cells become resistant to ·NO. Metallothionein (MT), the major protein thiol induced in cells exposed to cytokines and bacterial products, is capable of forming iron-dinitrosyl thiolates in vitro. Therefore, we tested the hypothesis that overexpression of MT reduces the sensitivity of NIH 3T3 cells to the ·NO donor, S-nitrosoacetylpenicillamine (SNAP), and to ·NO released from cells (NIH 3T3-DFG-iNOS) after infection with a retroviral vector expressing human iNOS gene. There was a 4-fold increase in MT in cells transfected with the mouse MT-I gene (NIH 3T3/MT) compared to cells transfected with the promoter-free inverted gene (NIH JT3/TM). NIH 3T3/MT cells were more resistant than NIH 3T3/TM cells to the cytotoxic effects of SNAP (0.1-1.0 mM) or ·NO released from NIH 3T3-DFG-iNOS cells. A brief (1 h) exposure to 10 mM SNAP caused DNA single-strand breaks that were 9-fold greater in NIH 3T3/TM compared to NIH 3T3/MT cells. Electron paramagnetic resonance spectroscopy of NIH 3T3 cells revealed a greater peak at g = 2.04 (e.g., iron-dinitrosyl complex) in NIH 3T3/MT than NIH 3T3/TM cells. These data are consistent with a role for cytoplasmic MT in interacting with ·NO and reducing ·NO-induced cyto- and nuclear toxicity.

Original languageEnglish
Pages (from-to)4452-4456
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number10
StatePublished - 9 May 1995
Externally publishedYes


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