Molecular and Cellular Biochemistry

, Volume 219, Issue 1–2, pp 29–37

Involvement of nuclear factor-I (NF1) binding motif in the regucalcin gene expression of rat kidney cortex: The expression is suppressed by cisplatin administration



The binding of nuclear factor on the promoter region of the regucalcin gene and the expression of regucalcin in the kidney cortex of rats was investigated. Nuclear extracts from kidney cortex were used for oligonucleotide competition gel mobility shift assay. An oligonucleotide between position –523 and –506 in the 5′-flanking region of the rat regucalcin gene, which contains a nuclear factor I (NF1) consensus motif TTGGC(N)6CC, competed with the probe for the binding of the nuclear protein from kidney cortex. The mutation of TTGGC in the consensus sequence caused an inhibition of the binding of nuclear factors. The binding of nuclear factor on the 5′-flanking region was clearly reduced in the kidney cortex obtained at 1, 2, and 3 days after a single intraperitoneal administration of cisplatin (1.0 mg/100 g body wt) to rats. Moreover, cisplatin administration caused a remarkable decrease in regucalcin mRNA levels and regucalcin concentration in the kidney cortex. Also, serum regucalcin concentration was significantly decreased by cisplatin administration. Meanwhile, serum urea nitrogen concentration was markedly elevated by cisplatin administration. The present study demonstrates that the specific nuclear factor binds to the NF1-like sequence in the promotor region of regucalcin gene in the kidney cortex of rats, and that the nuclear factor binding and regucalcin expression are suppressed by cisplatin administration.

regucalcin nuclear factor 1 (NF1) gene expression cisplatin rat kidney 


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  1. 1.
    Cheung WY: Calmodulin plays a pivotal role in cellular regulation. Science 202: 19–27, 1984Google Scholar
  2. 2.
    Nishizuka Y: Studies and perspectives of protein kinase C. Science 233: 305–312, 1986Google Scholar
  3. 3.
    Heizmann CW, Hunziker W: Intracellular calcium-binding proteins: More sites than in sight. Trends Biochem Sci 16: 98–103, 1991Google Scholar
  4. 4.
    Kraus-Friedman N, Feng L: The role of intracellular Ca2+ in the regulation of gluconeogenesis. Metabolism 45: 389–403, 1996Google Scholar
  5. 5.
    Yamaguchi M: Role of regucalcin in calcium signaling. Life Sci 66: 1769–1780, 2000Google Scholar
  6. 6.
    Yamaguchi M, Mori S: Inhibitory effect of calcium-binding protein regucalcin on protein kinase C activity in rat liver cytosol. Biochem Med Metab Biol 43: 140–146, 1990Google Scholar
  7. 7.
    Yamaguchi M, Katsumata T: Enhancement of protein kinase activity in the cytosol of regenerating rat liver: Regulatory role of endogenous regucalcin Int J Mol Med 3: 505–510, 1999Google Scholar
  8. 8.
    Kurota H, Yamaguchi M: Inhibitory effect of regucalcin on Ca2+/calmodulin-dependent protein kinase activity in rat renal cortex cytosol. Mol Cell Biochem 177: 239–243, 1997Google Scholar
  9. 9.
    Kurota H, Yamaguchi M: Inhibitory effect of calcium-binding protein regucalcin on protein kinase C activity in rat renal cortex cytosol. Biol Pharm Bull 21: 315–318, 1998Google Scholar
  10. 10.
    Shimokawa N, Matsuda Y, Yamaguchi M: Genomic cloning and chromosomal assignment of rat regucalcin gene. Mol Cell Biochem 151: 157–163, 1995Google Scholar
  11. 11.
    Yamaguchi M, Makino R, Shimokawa N: The 5′-end sequences and exon organization in rat regucalcin gene. Mol Cell Biochem 165: 145–150, 1996Google Scholar
  12. 12.
    Shimokawa N, Yamaguchi M: Calcium administration stimulates the expression of calcium-binding protein regucalcin mRNA in rat liver. FEBS Lett 305: 151–154, 1992Google Scholar
  13. 13.
    Shimokawa N, Yamaguchi M: Molecular cloning and sequencing of the cDNA coding for a calcium-binding protein regucalcin from rat liver. FEBS Lett 327: 251–255, 1993Google Scholar
  14. 14.
    Yamaguchi M, Isogai M: Tissue concentration of calcium-binding protein regucalcin in rats by enzyme-linked immunoadsorbent assay. Mol Cell Biochem 122: 65–68, 1993Google Scholar
  15. 15.
    Shimokawa N, Yamaguchi M: Expression of hepatic calcium-binding protein regucalcin mRNA mediated through Ca2+/calmodulin in rat liver. FEBS Lett 316: 79–84, 1993Google Scholar
  16. 16.
    Murata T, Yamaguchi M: Ca2+ administration stimulates the binding of AP-1 factor to the 5′-flanking region of the rat gene for the Ca2+-binding protein regucalcin. Biochem J 329: 157–163, 1998Google Scholar
  17. 17.
    Murata T, Yamaguchi M: Promoter characterization of the rat gene for Ca2+-binding protein regucalcin. Transcriptional regulation by signaling factors. J Biol Chem 274: 1277–1285, 1999Google Scholar
  18. 18.
    Yamaguchi M, Kurota H: Expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats: The stimulation by calcium administration. Mol Cell Biochem 146: 71–77, 1995Google Scholar
  19. 19.
    Kurota H, Yamaguchi M: Steroid hormonal regulation of calcium-binding protein regucalcin mRNA expression in the kidney cortex of rats. Mol Cell Biochem 155: 105–111, 1996Google Scholar
  20. 20.
    Shinya N, Kurota H, Yamaguchi M: Calcium-binding protein regucalcin mRNA expression in the kidney cortex is suppressed by saline ingestion in rats. Mol Cell Biochem 162: 139–144, 1996Google Scholar
  21. 21.
    Kurota H, Yamaguchi M: Suppressed expression of calcium-binding protein regucalcin mRNA in the renal cortex of rats with chemically induced kidney damage. Mol Cell Biochem 151: 55–60, 1995Google Scholar
  22. 22.
    Murata T, Yamaguchi M: Binding of kidney nuclear proteins to the 5′-flanking region of the rat gene for Ca2+-binding protein regucalcin: Involvement of Ca2+/calmodulin signaling. Mol Cell Biochem 199: 35–40, 1999Google Scholar
  23. 23.
    Dignam JD, Lebovitz RM, Roeder RG: Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 11: 1475–1489, 1983Google Scholar
  24. 24.
    Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976Google Scholar
  25. 25.
    Garner MM, Revzin A: A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: Application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res 9: 3047–3060, 1981Google Scholar
  26. 26.
    Yamaguchi M, Yamamoto T: Purification of calcium binding substance from soluble fraction of normal rat liver. Chem Pharm Bull 26: 1915–1918, 1978Google Scholar
  27. 27.
    Murata T, Yamaguchi M: Tissue-specific binding of nuclear factors to the 5′-flanking region of the rat gene for calcium-binding protein regucalcin. Mol Cell Biochem 178: 305–310, 1998Google Scholar
  28. 28.
    Heinemeyer T, Chen X, Karas H, Kel AE, Kel OV, Lieibich I, Meinhardt T, Reuter I, Schacherer F, Wingender E: Expanding the TRANSFAC database towards an expert system of regulatory molecular mechanisms. Nucleic Acids Res 27: 318–322, 1999Google Scholar
  29. 29.
    Krohn K, Rozovsky I, Wals P, Teter B, Anderson CP, Finch CE: Glial fibrillary acidic protein transcription responses to transforming growth factor-β1 and interleukin-1β are mediated by a nuclear factor-1-like site in the near-upstream promoter. J Neurochem 72: 1353–1361, 1999Google Scholar
  30. 30.
    Lee M, Song H, Park S, Park J: Transcription of the rat p53 gene is mediated by factor binding to two recognition motifs of NF1-like protein. Biol Chem 379: 1333–1340, 1998Google Scholar
  31. 31.
    Schuur ER, Kruse U, Iacovoni JS, Vogt PK: Nuclear factor I interferes with transformation induced by nuclear oncogenes. Cell Growth Diff 6: 219–227, 1995Google Scholar
  32. 32.
    Agus ZS, Chiu PJS, Goldberg M: Regulation of urinary calcium excretion in the rat. Am J Physiol 232: 545–549, 1997Google Scholar
  33. 33.
    Ng RCK, Peraino RA, Suki WN: Divalent cation transport in isolated tubules. Kidney Int 22: 492–497, 1982Google Scholar
  34. 34.
    Kurota H, Yamaguchi M: Activatory effect of calcium-binding protein regucalcin on ATP-dependent calcium transport in the basolateral membranes of rat kidney cortex. Mol Cell Biochem 169: 149–156, 1997Google Scholar
  35. 35.
    Ritzenthaler JD, Goldstein RH, Fine A, Smith BD: Regulation of the α1(I) collagen promoter via a transforming growth factor-beta activation element. J Biol Chem 268: 13625–13631, 1993Google Scholar
  36. 36.
    Ogata Y, Niisato N, Furuyama S, Cheifetz S, Kim RH, Sugiya H, Sodek J: Transforming growth factor-β1 regulation of bone sialoprotein gene transcription: identification of a TGF-β activation element in the rat BSP gene promoter. J Cell Biochem 65: 501–512, 1997Google Scholar
  37. 37.
    Montine TJ, Borch RF: Role of endogenous sulfur-containing nucleotides in an in vitro model of cis-diamminechloro platinum (II)-induced nephrotoxicity. Biochem Pharmacol 39: 1751–1757, 1990Google Scholar

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© Kluwer Academic Publishers 2001

Authors and Affiliations

  1. 1.Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional SciencesUniversity of ShizuokaShizuokaJapan

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