Activation of the growth arrest and DNA damage-inducible gene gadd 153 by nephrotoxic cysteine conjugates and dithiothreitol

Qin Chen, Kefu Yu, Nikki J. Holbrook, James L. Stevens

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

The cellular and biochemical events which transduce chemical insults into signals for increased expression of the stress-responsive gene gadd 153 were investigated using nephrotoxic cysteine conjugates. In LLC-PK1 cells, cysteine conjugate toxicity is initiated by covalent binding, but depletion of cellular thiols, an increase in cytosolic free calcium, and lipid peroxidation couple the binding to cell death (Chen, Q., Jones, T. W., Brown, P. C., and Stevens, J. L. (1990) J. Biol. Chem. 265, 21603-21611; Chen, Q., Jones, T. W., and Stevens, J. L. (1991) Toxicologist 11, 101, 1991). Three different toxic cysteine conjugates induced gadd 153 mRNA. With S-(1,2-dichlorovinyl)-L-cysteine (DCVC), the induction was both concentration and time-dependent. Preventing the metabolism of DCVC and covalent binding of DCVC-derived reactive metabolites to cellular macromolecules with the β-lyase inhibitor (aminooxy)acetic acid blocked the induction. However, buffering free calcium with a cell permeable calcium chelator or blocking lipid peroxidation with an antioxidant did not affect the induction of gadd 153 mRNA by DCVC even though these treatments inhibit toxicity. These data suggest that covalent binding of reactive metabolites to cellular macromolecules may serve as a primary signal for the induction of gadd 153 mRNA by nephrotoxic cysteine conjugates. Interestingly, the sulfhydryl agent dithiothreitol, which was nontoxic and prevented the toxicity of DCVC, also induced an increase in gadd 153 mRNA. When both dithiothreitol and DCVC were added to cells, there were no inhibitory or additive effects on expression. Therefore, cellular thiol-disulfide status may also play a role in gadd 153 induction.

Original languageEnglish (US)
Pages (from-to)8207-8212
Number of pages6
JournalJournal of Biological Chemistry
Volume267
Issue number12
StatePublished - Apr 25 1992
Externally publishedYes

Fingerprint

Dithiothreitol
DNA Damage
Cysteine
Genes
Chemical activation
DNA
Growth
Toxicity
Messenger RNA
Metabolites
Macromolecules
Sulfhydryl Compounds
Lipid Peroxidation
Aminooxyacetic Acid
Calcium
Lipids
LLC-PK1 Cells
Lyases
Poisons
Cell death

ASJC Scopus subject areas

  • Biochemistry

Cite this

Activation of the growth arrest and DNA damage-inducible gene gadd 153 by nephrotoxic cysteine conjugates and dithiothreitol. / Chen, Qin; Yu, Kefu; Holbrook, Nikki J.; Stevens, James L.

In: Journal of Biological Chemistry, Vol. 267, No. 12, 25.04.1992, p. 8207-8212.

Research output: Contribution to journalArticle

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