Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Calmodulin binds to a tubulin binding site of the microtubule-associated protein tau

  • 114 Accesses

  • 34 Citations

Abstract

Previous studies have demonstrated that the microtubule - associated proteins MAP-2 and tau interact selectively with common binding domains on tubulin defined by the low-homology segments a (430–441) and β (422–434). It has been also indicated that the synthetic peptide VRSKIGSTENLKHQPGGG corresponding to the first tau repetitive sequence represents a tubulin binding domain on tau. The present studies show that the calcium-binding protein calmodulin interacts with a tubulin binding site on tau defined by the second repetitive sequence VTSKCGSLGNIHHKPGGG. It was shown that both tubulin and calmodulin bind to tau peptide-Sepharose affinity column. Binding of calmodulin occurs in the presence of 1 mM Ca 2+ and it can be eluted from the column with 4 mM EGTA. These findings provide new insights into the regulation of microtubule assembly, since Ca 2+/calmodulin inhibition of tubulin polymerization into microtubules could be mediated by the direct binding of calmodulin to tau, thus preventing the interaction of this latter protein with tubulin.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Weisenberg RC: Mucrotubule formation in vitro in solutions containing calcium concentrations. Science 177: 1104–1105,1972

  2. 2.

    Berkowitz SA, Wolff JC: Intrinsic calcium sensitivity of tubulin polymerization. J Biol Chem 257: 11216–11223, 1981

  3. 3.

    Solomon F: Binding sites for calcium and calmodulin. Biochemistry 16: 358–363, 1977

  4. 4.

    Serrano L, Valencia A, Caballero R, Avila J: Localization of the high affinity calcium binding site on tubulin molecule. J Biol Chem 261: 7076–7081, 1986

  5. 5.

    Marcum JM, Dedman JR, Brinkley BR, Means AR: Control of microtubule assembly-disassembly by calcium dependent regulator proteins. Proc Natl Acad Sci USA 75: 3771–3775, 1978

  6. 6.

    Job D, Fisher EH, Margolis RC: Rapid disassembly of cold-stable microtubules by calmodulin. Proc Natl Acad Sci USA 78: 4679–4682, 1981

  7. 7.

    Lee YC, Wolff J: Two opposing effects of calmodulin of microtubule assembly depend on the presence of microtubule-associated proteins. J Biol Chem 257: 6302–6310, 1982

  8. 8.

    Keller TCS, Jemiolo DC, Burgess WH, Rebhum LI: Strongylocentrotus purpuratus spindle tubulin. II. Characteristics of its sensitivity to Ca 2 + and the effects of calmodulin isolated from bovine brain and S. purpuratus. J Cell Biol 93: 797–803, 1982

  9. 9.

    Rebhum LI, Jemiolo D, Keller T, Burgess W, Ketsinger R: In: M De Brabander and M De May (eds) Microtubules and Microtubule Inhibitors. Elsevier-North Holland Press, Amsterdam, 1983, pp. 243–252

  10. 10.

    Erneux C, Passareiro H, Nunez J: Interaction between calmodulin and microtubule-associated proteins prepared at different stages of brain development. FEBS Lett 172: 315–320, 1984

  11. 11.

    Bender PK, Rebhum LI: The calcium sensitivity of MAP-2 and Tau microtubules in the presence of calmodulin. Ann NY Acad Sci 466: 392–409, 1986

  12. 12.

    Lee YC, Wolff J: Calmodulin binds to both microtubule associated protein 2 and tau proteins. J Biol Chem 259: 1226–1230, 1984

  13. 13.

    Jemiolo DK, Burgess WH, Rebhum LI, Kretsinger RH: Calmodulin interaction with cycle-purified brain tubulin components. J Cell Biol 87: 248a, 1980

  14. 14.

    Kumagai H, Nishida E, Kotani S, Sakai H: On the mechanism of calmodulin-induced inhibition of microtubule assembly in vitro. J Biochem 99: 521–525, 1986

  15. 15.

    Sobue K, Fujita M, Muramoto K, Kariuchi S: The calmodulin binding protein in microtubules is Tau factor. FEBS Lett 137–140, 1981

  16. 16.

    Lee YC, Wolff J: The calmodulin binding domain on microtubule-associated protein 2. J Biol Chem 259: 8041–8044, 1984

  17. 17.

    Lee G, Cowan N, Kirschner MW: The primary structure and heterogeneity of tau protein from mouse brain. Science 239:285–288,1988

  18. 18.

    Lewis SA, Wang D, Cowan NJ: Microtubule associated protein MAP-2 shares a microtubule binding motif with tau protein. Science 242: 936–939, 1988

  19. 19.

    Goedert M, Wischik CM, Crowther RA, Walker JE, Klug A: Cloning and sequencing of the cDNA encoding a core protein of the paired helical filament of Alzheimer disease: Identification as microtubule-associated protein tau. Proc Natl Acad Sci USA 85: 4051–4055, 1988

  20. 20.

    Maccioni RB, Rivas CI, Vera JC: Differential interaction of synthetic peptides from the carboxy-terminal regulatory domain of tubulin with microtubule associated proteins (MAPS). EMBO J 7: 1057–1963, 1988

  21. 21.

    Shelanski LM, Gaskin F, Cantor CR: Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci USA 70: 765–768, 1973

  22. 22.

    Weingarten MD, Lockwood AH, Hwo SY, Kirschner MW: A protein factor essential for microtubule assembly. Proc Natl Acad Sci USA 72: 1858–1862, 1975

  23. 23.

    Coyne MD, Cornelius P, Venditti N, Toscano DG, Gross MK, Tossano WA: Purification and properties of calmodulin from adrenal cortex. Arch Biochem Biophys 236: 629–637, 1985

  24. 24.

    Laemmli UK: Cleavage of structural proteins during the assembly of head of bacteriophage. Nature 227: 680–685, 1970

  25. 25.

    Vera JC, Rivas C, Maccioni RB: Biochemical dissection of the role of the 1 kDA C-terminal domain of tubulin subunits in the assembly of microtubules. Biochemistry 28: 333–339, 1989

  26. 26.

    Mitchison T, Kirschner M: Microtubule assembly nucleated by isolated centrosomes. Nature 312: 232–237, 1984

  27. 27.

    Maccioni RB, Vera JC, Dominguez J, Avila J: A discrete repeated sequence defines a tubulin binding domain on tau. Arch Biochem Biophys 275: 568–579, 1989

  28. 28.

    Baudier J: Comparison of S100b protein with calmodulin: Interaction with mellitin and microtubule associated tau proteins and inhibition of phosphorylation of tau proteins by protein kinase C. Biochemistry 26: 2886–2893, 1987

  29. 29.

    Serrano J, Avila J, Maccioni RB: Controlled proteolysis of tubulin by subtilisin: localization of the site for MAP-2 interaction. Biochemistry 23: 4675–4681, 1984

  30. 30.

    Serrano L, de la Torre J, Maccioni RB, Avila J: Involvement of the carboxyl-terminal domain of tubulin in microtubule assembly and regulation. Proc Natl Acad Sci USA 81: 5989–5993, 1984

  31. 31.

    Littauer UZ, Giveon D, Thierauf M, Ginzburg I, Ponstingl H: Common and distinct tubulin binding sites for microtubule associated proteins. Proc Natl Acad Sci USA 83: 7162–7166, 1986

  32. 32.

    Littauer UZ, Giveon D, Thierauf M, Ginzburg I, Ponstingl H: In: M De Brabander and M De May (eds) Microtubules and Microtubule Inhibitors, Elsevier North Holland Biomedical Press, 1985; pp 171–176

  33. 33.

    Dedman JR, Welsch MJ, Means AR: Ca 2 + dependent regulator: Production and characterization of a monospecific antibody. J Biol Chem 253: 7515–7521, 1978

  34. 34.

    Watterson DM, Mendel PA, Vanaman TC: Comparison of calcium-modulated proteins from vertebrate brains. Biochemistry 19: 2672–2676, 1980

  35. 35.

    Baudier J, Cole RD: Interaction between the microtubule associated tau protein and 100b regulates tau phosphorylation by the Ca2 +/calmodulin-dependent protein kinase II. J Biol Chem 263: 5876–5883, 1988

  36. 36.

    Kakiuchi S, Sobue K: Calcium and calmodulin flip-flop mechanism in microtubule assembly-disassembly. FEBS Lett 132: 141–143, 1981

  37. 37.

    Ponstingl H, Krauhs E, Little M, Kempf M, Hofer Warbinek R, Ade W: Amino acid sequence of α and β tubulins from pig brain: heterogeneity and regional similarity to muscle proteins. Cold Spring Harbor Symp Quant Biol 46: 191–198, 1982

  38. 38.

    Isobe T, Akuyama T: The amino acid sequence of the α-subunit in bovine brain S-100a protein. Eur J Biochem 116: 76–86, 1981

Download references

Author information

Correspondence to Ricardo B. Maccioni.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Padilla, R., Maccioni, R.B. & Avila, J. Calmodulin binds to a tubulin binding site of the microtubule-associated protein tau. Mol Cell Biochem 97, 35–41 (1990). https://doi.org/10.1007/BF00231699

Download citation

Key words

  • tau protein
  • tubulin binding site
  • calmodulin interaction
  • microtubule assembly