Signalling the molecular stress response to nephrotoxic and mutagenic cysteine conjugates: Differential roles for protein synthesis and calcium in the induction of c‐fos and c‐myc mRNA in LLC‐PK1 cells

Nephrotoxic and mutagenic cysteine conjugates (NCC) are activated by the enzyme cysteine conjugate, β‐lyase, to reactive acylating species which bind covalently to cellular macromolecules. We now show that an early event after treatment of LLC‐PK1 cells with NCC is the induction of mRNA for both c‐fos and c‐myc. Treatment with S‐(1,2‐dichlorovinyl)‐L‐cysteine (DCVC) induced c‐fos (53‐fold) and c‐myc mRNA (20‐fold) and increased transcription about 3‐fold for both genes. Covalent binding was required for induction of both mRNAs. Dithiothreitol partially prevented induction of both c‐fos and c‐myc RNA. Buffering the DCVC‐induced increase in cytosolic free calcium had no effect on c‐fos mRNA but partially blocked c‐myc mRNA induction. Cycloheximide blocked the induction of c‐myc mRNA in the absence of an effect on c‐fos induction. The data suggest that the increase in c‐fos mRNA is a primary response to DCVC toxicity and occurs without a requirement for protein synthesis or an increase in intracellular free calcium. In contrast, c‐myc induction requires protein synthesis, suggesting that the presence of another primary response factor may regulate induction either transcriptionally or posttranscriptionally. The data suggest that different signalling pathways regulate induction of c‐fos and c‐myc mRNA in response to stress caused by reactive acylating species. © 1994 Wiley‐Liss, Inc.

[1]  L. Cantley,et al.  Oncogenes and signal transduction , 1991, Cell.

[2]  D. Hallahan,et al.  Ionizing radiation regulates expression of the c-jun protooncogene. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[3]  M. W. Anders,et al.  Perturbations of intracellular calcium distribution in kidney cells by nephrotoxic haloalkenyl cysteine S-conjugates. , 1990, Molecular pharmacology.

[4]  K. Nose,et al.  Stimulation by hydrogen peroxide of DNA synthesis, competence family gene expression and phosphorylation of a specific protein in quiescent Balb/3T3 cells. , 1990, Oncogene.

[5]  P. Bernstein,et al.  Poly(A), poly(A) binding protein and the regulation of mRNA stability. , 1989, Trends in biochemical sciences.

[6]  G. Storz,et al.  Spontaneous mutagenesis and oxidative damage to DNA in Salmonella typhimurium. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[7]  N. Holbrook,et al.  Calcium ionophore A23187 induces expression of the growth arrest and DNA damage inducible CCAAT/enhancer-binding protein (C/EBP)-related gene, gadd153. Ca2+ increases transcriptional activity and mRNA stability. , 1992, The Journal of biological chemistry.

[8]  Q. Chen,et al.  Inhibition of iodoacetamide and t-butylhydroperoxide toxicity in LLC-PK1 cells by antioxidants: a role for lipid peroxidation in alkylation induced cytotoxicity. , 1991, Archives of biochemistry and biophysics.

[9]  B van de Water,et al.  Role of mitochondrial Ca2+ in the oxidative stress-induced dissipation of the mitochondrial membrane potential. Studies in isolated proximal tubular cells using the nephrotoxin 1,2-dichlorovinyl-L-cysteine. , 1994, The Journal of biological chemistry.

[10]  R. Morimoto,et al.  Structure and expression of the human gene encoding major heat shock protein HSP70 , 1985, Molecular and cellular biology.

[11]  Q. Chen,et al.  Regulation of the cellular stress response by reactive electrophiles. The role of covalent binding and cellular thiols in transcriptional activation of the 70-kilodalton heat shock protein gene by nephrotoxic cysteine conjugates. , 1992, The Journal of biological chemistry.

[12]  E. Lock,et al.  Pentachlorobutadienyl-L-cysteine (PCBC) toxicity: the importance of mitochondrial dysfunction. , 1991, Journal of biochemical toxicology.

[13]  J. Harris,et al.  In vivo detection and characterization of protein adducts resulting from bioactivation of haloethene cysteine S-conjugates by 19F NMR: chlorotrifluoroethene and tetrafluoroethene. , 1992, Chemical research in toxicology.

[14]  J. Stevens,et al.  Early cellular events couple covalent binding of reactive metabolites to cell killing by nephrotoxic cysteine conjugates , 1994, Journal of cellular physiology.

[15]  I. Verma,et al.  Nuclear proto-oncogenes fos and jun. , 1990, Annual review of cell biology.

[16]  J. Stevens,et al.  Cysteine conjugate toxicity, metabolism, and binding to macromolecules in isolated rat kidney mitochondria. , 1990, Molecular pharmacology.

[17]  G. Stein,et al.  Disruption of the cytoskeleton with cytochalasin D induces c-fos gene expression. , 1991, Experimental cell research.

[18]  S. Farr,et al.  Oxidative stress responses in Escherichia coli and Salmonella typhimurium. , 1991, Microbiological reviews.

[19]  S. Vamvakas,et al.  Enhanced expression of the protooncogenes c-myc and c-fos in normal and malignant renal growth. , 1993, Toxicology letters.

[20]  T. Curran,et al.  Identification and characterization of Ref‐1, a nuclear protein that facilitates AP‐1 DNA‐binding activity. , 1992, The EMBO journal.

[21]  J. Farber,et al.  The role of calcium in lethal cell injury. , 1990, Chemical research in toxicology.

[22]  A. Fornace,et al.  Response to adversity: molecular control of gene activation following genotoxic stress. , 1991, The New biologist.

[23]  J. Calvet,et al.  The heat shock response in HeLa cells is accompanied by elevated expression of the c-fos proto-oncogene , 1987, Molecular and cellular biology.

[24]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[25]  P. Hayden,et al.  Cysteine S-conjugates. , 1987, Methods in enzymology.

[26]  J. Stevens,et al.  The role of glutathione conjugate metabolism and cysteine conjugate beta-lyase in the mechanism of S-cysteine conjugate toxicity in LLC-PK1 cells. , 1986, The Journal of biological chemistry.

[27]  D. Ron,et al.  CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcription factors C/EBP and LAP and functions as a dominant-negative inhibitor of gene transcription. , 1992, Genes & development.

[28]  J. Stevens,et al.  The mechanism of cysteine conjugate cytotoxicity in renal epithelial cells. Covalent binding leads to thiol depletion and lipid peroxidation. , 1990, The Journal of biological chemistry.

[29]  S. Orrenius,et al.  The mechanism of pentachlorobutadienyl-glutathione nephrotoxicity studied with isolated rat renal epithelial cells. , 1986, Archives of biochemistry and biophysics.

[30]  D. Hallahan,et al.  Protein kinase C mediates x-ray inducibility of nuclear signal transducers EGR1 and JUN. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[31]  D. J. Reed,et al.  Review of the current status of calcium and thiols in cellular injury. , 1990, Chemical research in toxicology.

[32]  D. Hallahan,et al.  Involvement of reactive oxygen intermediates in the induction of c-jun gene transcription by ionizing radiation. , 1992, Biochemistry.

[33]  J. Fargnoli,et al.  Role of [Ca2+]i in induction of c‐fos, c‐jun and c‐myc mRNA in rat PTE after oxidative stress , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[34]  T. Curran,et al.  FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA , 1982, Journal of virology.

[35]  W. H. Schaefer,et al.  Formation of mitochondrial phospholipid adducts by nephrotoxic cysteine conjugate metabolites. , 1992, Chemical research in toxicology.

[36]  N. Vermeulen,et al.  Molecular and biochemical mechanisms of chemically induced nephrotoxicity: a review. , 1990, Chemical research in toxicology.

[37]  W. Dekant,et al.  Events that precede and that follow S-(1,2-dichlorovinyl)-L-cysteine-induced release of mitochondrial Ca2+ and their association with cytotoxicity to renal cells. , 1992, Biochemical pharmacology.

[38]  M. Karin,et al.  Activation of the c-fos gene by UV and phorbol ester: different signal transduction pathways converge to the same enhancer element. , 1988, Oncogene.

[39]  R. Byrd,et al.  A purified cysteine conjugate beta-lyase from rat kidney cytosol. Requirement for an alpha-keto acid or an amino acid oxidase for activity and identity with soluble glutamine transaminase K. , 1986, The Journal of biological chemistry.

[40]  I. Verma,et al.  Regulated mRNA stability. , 1990, Annual review of genetics.

[41]  S. Orrenius,et al.  The role of Ca2+ in cell killing. , 1990, Chemical research in toxicology.

[42]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[43]  C. Gross,et al.  8 The Function and Regulation of Heat Shock Proteins in Escherichia coli , 1990 .

[44]  Richard J Smeyne,et al.  Continuous c-fos expression precedes programmed cell death in vivo , 1993, Nature.

[45]  T. Curran,et al.  Redox activation of Fos‐Jun DNA binding activity is mediated by a DNA repair enzyme. , 1992, The EMBO journal.

[46]  H. Herschman.  Primary response genes induced by growth factors and tumor promoters. , 1991, Annual review of biochemistry.

[47]  N. Holbrook,et al.  Activation of the growth arrest and DNA damage-inducible gene gadd 153 by nephrotoxic cysteine conjugates and dithiothreitol. , 1992, The Journal of biological chemistry.

[48]  G C Walker,et al.  Inducible DNA repair systems. , 1985, Annual review of biochemistry.

[49]  S. Orrenius,et al.  Calcium-activated DNA fragmentation in rat liver nuclei. , 1989, The Journal of biological chemistry.

[50]  F. Herrmann,et al.  Activation of the transcription factor kappa B in human KG-1 myeloid leukemia cells treated with 1-beta-D-arabinofuranosylcytosine. , 1992, Molecular pharmacology.

[51]  M. Brostrom,et al.  Accommodation of protein synthesis to chronic deprivation of intracellular sequestered calcium. A putative role for GRP78. , 1990, The Journal of biological chemistry.

[52]  R. Morimoto,et al.  1 The Stress Response, Function of the Proteins, and Perspectives , 1990 .

[53]  J. Stevens,et al.  Formation of difluorothionoacetyl-protein adducts by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine metabolites: nucleophilic catalysis of stable lysyl adduct formation by histidine and tyrosine. , 1991, Biochemistry.

[54]  M. Karin,et al.  The mammalian ultraviolet response is triggered by activation of src tyrosine kinases , 1992, Cell.

[55]  R. Morimoto,et al.  Transcription of the Human HSP70 Gene: cis-Acting Elements and trans-Acting Factors Involved in Basal, Adenovirus E1A, and Stress-Induced Expression , 1990 .

[56]  P. Baeuerle,et al.  Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF‐kappa B transcription factor and HIV‐1. , 1991, The EMBO journal.

[57]  A. Mantovani,et al.  Heat shock induces the transcriptional activation of c-fos protooncogene. , 1990, Biochemical and biophysical research communications.

[58]  M. Karin,et al.  Rapid and preferential activation of the c-jun gene during the mammalian UV response , 1991, Molecular and cellular biology.

[59]  S. Orrenius,et al.  Activation of hepatocyte protein kinase C by redox-cycling quinones. , 1989, The Biochemical journal.

[60]  E. Lock,et al.  The effects of haloalkene cysteine conjugates on cytosolic free calcium levels in suspensions of rat renal proximal tubules. , 1990, Journal of biochemical toxicology.

[61]  R. Schnellmann.  Intracellular calcium chelators and oxidant-induced renal proximal tubule cell death. , 1991, Journal of biochemical toxicology.

[62]  W. Dekant,et al.  Bioactivation of nephrotoxic haloalkenes by glutathione conjugation: formation of toxic and mutagenic intermediates by cysteine conjugate beta-lyase. , 1989, Drug metabolism reviews.

[63]  B. van de Water,et al.  The relationship between intracellular Ca2+ and the mitochondrial membrane potential in isolated proximal tubular cells from rat kidney exposed to the nephrotoxin 1,2-dichlorovinyl-cysteine. , 1993, Biochemical pharmacology.

[64]  D. Wolgemuth,et al.  Cascade induction of c-fos, c-myc, and heat shock 70K transcripts during regression of the rat ventral prostate gland. , 1988, Molecular endocrinology.

[65]  M. Freedman,et al.  The expression of c-fos, c-jun, and c-myc genes is regulated by heat shock in human lymphoid cells. , 1990, Journal of immunology.

[66]  N. Heintz,et al.  Persistent induction of c-fos and c-jun expression by asbestos. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[67]  P. Herrlich,et al.  UV-induced DNA damage is an intermediate step in UV-induced expression of human immunodeficiency virus type 1, collagenase, c-fos, and metallothionein , 1989, Molecular and cellular biology.

[68]  W. Anderson,et al.  Ca2+- and phospholipid-independent activation of protein kinase C by selective oxidative modification of the regulatory domain. , 1989, Proceedings of the National Academy of Sciences of the United States of America.