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Molecular and Cellular Biochemistry

, Volume 199, Issue 1–2, pp 35–40 | Cite as

Binding of kidney nuclear proteins to the 5′-flanking region of the rat gene for Ca2+-binding protein regucalcin: Involvement of Ca2+/calmodulin signaling

  • Tomiyasu Murata
  • Masayoshi Yamaguchi
Article

Abstract

Ca2+-binding protein regucalcin is expressed in the kidney cortex of rats, as assayed by Northern blot analysis. The existence of kidney nuclear factor which binds to the 5'-flanking region of the rat regucalcin gene was investigated. When nuclear extracts obtained from the kidney cortex of rats were used in gel mobility-shift assays, two protein-DNA complexes were uniquely formed with the DNA fragment containing the 5'-flanking region of the rat regucalcin gene. Competition gel shift experiments indicated the specific binding region of kidney cortex nuclear proteins in the 5′-flanking region of the rat regucalcin gene. The two nuclear protein-DNA complexes were formed with the same mobility in rat kidney cortex and liver, which possess detectable amounts of regucalcin mRNA in Northern blot analysis. The binding activities of nuclear factors from kidney cortex to the 5′-flanking region of the rat regucalcin gene were inhibited by a single intraperitoneal administration of trifluoperazine, an antagonist of calmodulin, to rats. The present study demonstrates that kidney cortex nuclear proteins specifically bind to the 5′-flanking region of the rat regucalcin gene, and that the binding activity may be partly mediated through the Ca2+/calmodulin-dependent process.

regucalcin gene nuclear factor Ca2+/calmodulin Gel mobility-shift assay rat kidney cortex 

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References

  1. 1.
    Wasserman RH: Calcium-binding proteins: Past, present and future. In: C.W. Heizmann (ed). Novel Calcium-Binding Proteins. Springer-Verlag, Berlin, 1991, pp 7–12Google Scholar
  2. 2.
    Yamaguchi M: A novel Ca2+-binding protein regucalcin and calcium inhibition: Regulatory role in liver cell function. In: K. Kohama (ed). Calcium Inhibition. Japan Sci Soc Press, Tokyo and CRC Press, Boca Raton, 1992, pp 19–41Google Scholar
  3. 3.
    Yamaguchi M, Tai H: Inhibitory effect of calcium-binding protein regucalcin on Ca2+/calmodulin-dependent cyclic nucleotide phosphodiesterase activity in rat liver cytosol. Mol Cell Biochem 106: 25–301 1991Google Scholar
  4. 4.
    Yamaguchi M, Sakurai T: Inhibitory effect of calcium-binding protein regucalcin on Ca2+-activated DNA fragmentation in rat liver nuclei. FEBS Lett 279: 281–284, 1991Google Scholar
  5. 5.
    Shimokawa N, Matsuda Y, Yamaguchi M: Genomic cloning and chromosomal assignment of rat regucalcin gene. Mol Cell Biochem 151: 157–163, 1995Google Scholar
  6. 6.
    Shimokawa N, Isogai M, Yamaguchi M: Specific species and tissue differences for the gene expression of calcium-binding protein regucalcin. Mol Cell Biochem 143: 67–71, 1995Google Scholar
  7. 7.
    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
  8. 8.
    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
  9. 9.
    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
  10. 10.
    Kurota H, Yamaguchi M: Steroid hormonal regulation of calciumbinding protein regucalcin mRNA expression in the kidney cortex of rats. Mol Cell Biochem 155: 105–111, 1996Google Scholar
  11. 11.
    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
  12. 12.
    Tamai Y, Silos SA, Li Y, Korkeela E, Ishikawa H, Uitto J: Tissuespecific expression of the 230-kDa bullous pemphigoid antigen gene (BPAG1). Identification of a novel keratinocyte regulatory cis-element KRE3. J Biol Chem 270: 7609–7614, 1995Google Scholar
  13. 13.
    Schmoelzl S, Leeb T, Brinkmeier H, Brem G, Brenig B: Regulation of tissue-specific expression of the skeletal muscle ryanodine receptor gene. J Biol Chem 271: 4763–4769, 1996Google Scholar
  14. 14.
    Brenner R, Thomas TO, Becker MN, Atkinson NS: Tissue-specific expression of a Ca2+-activated K+ channel is controlled by multiple upstream regulatory elements. J Neurosci 16: 1827–1835, 1996Google Scholar
  15. 15.
    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
  16. 16.
    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
  17. 17.
    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: 3 047–3060, 1981Google Scholar
  18. 18.
    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
  19. 19.
    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
  20. 20.
    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
  21. 21.
    Weiss B, Wallace TL: Mechanisms and pharmacological implications of altering calmodulin activity. In: W.Y. Cheung (ed). Calcium and Cell Function, Vol. 1. Academic Press, New York, 1980, pp 329–379Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Tomiyasu Murata
    • 1
  • Masayoshi Yamaguchi
    • 1
  1. 1.Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional SciencesUniversity of ShizuokaShizuoka CityJapan

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