Journal of Molecular Evolution

, Volume 43, Issue 4, pp 384–398 | Cite as

Molecular evolution of the 14-3-3 protein family

  • Wenfu Wang
  • Diane C. Shakes


Members of the highly conserved and ubiquitous 14-3-3 protein family modulate a wide variety of cellular processes. To determine the evolutionary relationships among specific 14-3-3 proteins in different plant, animal, and fungal species and to initiate a predictive analysis of isoform-specific differences in light of the latest functional and structural studies of 14-3-3, multiple alignments were constructed from forty-six 14-3-3 sequences retrieved from the GenBank and SwissProt databases and a newly identified second 14-3-3 gene fromCaenorhabditis elegans. The alignment revealed five highly conserved sequence blocks. Blocks 2–5 correlate well with the alpha helices 3, 5, 7, and 9 which form the proposed internal binding domain in the three-dimensional structure model of the functioning dimer. Amino acid differences within the functional and structural domains of plant and animal 14-3-3 proteins were identified which may account for functional diversity amongst isoforms. Protein phylogenic trees were constructed using both the maximum parsimony and neighbor joining methods of the PHYLIP(3.5c) package; 14-3-3 proteins fromEntamoeba histolytica, an amitochondrial protozoa, were employed as an outgroup in our analysis. Epsilon isoforms from the animal lineage form a distinct grouping in both trees, which suggests an early divergence from the other animal isoforms. Epsilons were found to be more similar to yeast and plant isoforms than other animal isoforms at numerous amino acid positions, and thus epsilon may have retained functional characteristics of the ancestral protein. The known invertebrate proteins group with the nonepsilon mammalian isoforms. Most of the current 14-3-3 isoform diversity probably arose through independent duplication events after the divergence of the major eukaryotic kingdoms. Divergence of the seven mammalian isoforms beta, zeta, gamma, eta, epsilon, tau, and sigma (stratifin/ HME1) occurred before the divergence of mammalian and perhaps before the divergence of vertebrate species. A possible ancestral 14-3-3 sequence is proposed.

Key words

14-3-3 Protein family Phylogenic tree Isoform diversity C. elegans 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alam R, Hachiya N, Sakaguchi M, Kawabata S, Iwanaga S, Kitajima M, Mihara K, Omura T (1994) cDNA cloning and characterization of mitochondrial import stimulation factor (MSF) purified from rat liver cytosol. J Biochem (Tokyo) 1116(2): 416–425Google Scholar
  2. Aitken A (1995) 14-3-3 proteins on the MAP. Trends Biochem Sci 20(3): 95–97CrossRefPubMedGoogle Scholar
  3. Aitken A, Collinge DB, van Heusden PH, Isobe T, Roseboom PH, Rosenfeld G, Soll J (1992) 14-3-3 proteins: a highly conserved widespread family of eukaryotc proteins. Trends Biochem Sci 17: 498–501CrossRefPubMedGoogle Scholar
  4. Aitken A, Howell S, Jones D, Madrazo J, Patel Y (1995) 14-3-3 alpha and delta are the phosphorylated forms of raf-activating 14-3-3 beta and zeta.In vivo stoichiometric phosphorylation in brain at a Ser-Pro-Glu-Lys motif. J Biol Chem 17, 270(11): 5706–5709Google Scholar
  5. Bairoch A, Bucher P (1994) Prosite: recent developments. Nucleic Acids Res 22: 3583–3589PubMedGoogle Scholar
  6. Baldauf SL, Palmer JD (1993) Animals and fungi are each other's closest relatives: congruent evidence from multiple proteins. PNAS 90: 11558–11562PubMedGoogle Scholar
  7. Brandt J, Thordalchristensen H, Vad K, Gregersen PL, Collinge DB (1992) A pathogen-induced gene of barley encodes a protein showing high similarity to a protein-kinase regulator. Plant J 2 (5):815–820CrossRefPubMedGoogle Scholar
  8. Burbelo PD, Hall A (1995) 14-3-3 proteins. Hot numbers in signal transduction. Curr Biol 5 (2): 95–96CrossRefPubMedGoogle Scholar
  9. Cavalier-Smith T, Allsopp MTEP, Chao EE (1994) Thraustochytrids are chromists, not Fungi: 18s rRNA signatures of Heterokonta. Philos Trans R Soc Lond Biol 346: 387–397Google Scholar
  10. Conklin DS, Galaktionov K, Beach D (1995) 14-3-3 proteins associate with cdc25 phosphatases. PNAS 92: 7892–7896PubMedGoogle Scholar
  11. deVetten NC, Lu GH, Ferl RJ (1992) A maize pattern associated with the G-box binding complex has homology to brain regulatory proteins. Plant Cell 4 (10): 1295–1307Google Scholar
  12. De Rijk P, Van de Peer Y, Van den Broeck I, De Wachter R (1995) Evolution according to large ribosomal subunit RNA. J Mol Evol 41: 366–375PubMedGoogle Scholar
  13. Doolittle RF (1993) Convergent evolution: the need to be concise. Trends Biochem Sci 19: 15–18Google Scholar
  14. Fantl WJ, Muslin AJ, Kikuchi A, Martin JA, MacNicol AM, Gross R, Williams LT (1994) Activation of Raf-1 by 14-3-3 proteins. Nature 13, 371 (6498): 612–614Google Scholar
  15. Felsenstein J (1993) PHYLIP(phylogeny inference package) version 3.5c. Distributed by the author. Department of Genetics, University of Washington, SeattleGoogle Scholar
  16. Ferl RJ, Lu G, Bowen BW (1994) Evolutionary implications of the family of 14-3-3 brain protein homologs inArabidopsis thaliana. Genetica 92(2): 129–138CrossRefPubMedGoogle Scholar
  17. Ford JC, al-Khodairy F, Fotou E, Sheldrick KS, Griffiths DJ, Carr AM (1994) 14-3-3 protein homologs required for the DNA damage checkpoint in fission yeast. Science 265: 533–535PubMedGoogle Scholar
  18. Freed E, Symons M, Macdonald SG, McCormick F, Ruggieri R (1994) Binding of 14-3-3 proteins to the protein kinase Raf and effects on its activation. Science 265: 1713–1716PubMedGoogle Scholar
  19. Fu H, Coburn J, Collier RJ (1993) The eukaryotic host factor that activates exoenzyme S ofPseudomonas aeruginosa is a member of the 14-3-3 protein family. PNAS 90: 2320–2324PubMedGoogle Scholar
  20. Fu H, Xia K, Pallas DC, Cui C, Conroy K, Narsimhan RP, Mamon H, Collier RJ, Roberts TM (1994) Interaction of the protein kinase Raf-1 with 14-3-3 proteins. Science 266: 126–129PubMedGoogle Scholar
  21. Hasegawa M, Hashimoto T, Adachi J, Iwabe N, Miyata T (1993) Early branchings in the evolution of eukaryotes: ancient divergence of Entamoeba that lacks mitochondria revealed by protein sequence data. J Mol Evol 36: 380–388CrossRefPubMedGoogle Scholar
  22. Hinkle G, Leipe DD, Nerad TA, Sogin ML (1994) The unusually long small subunit ribosomal RNA ofPhreatamoeba balamuthi. Nucleic Acids Res 22: 465–469PubMedGoogle Scholar
  23. Hirsch S, Aitken A, Bertsch U, Soll J (1992) A plant homologue to mammalian brain 14-3-3 protein and protein kinase C inhibitor. FEBS Lett 296: 222–224CrossRefPubMedGoogle Scholar
  24. Ichimura T, Isobe T, Okuyama T, Takahashi N, Araki K, Kuwano R, Takahashi Y (1988) Molecular cloning of cDNA coding for brain-specific 14-3-3 protein, a protein kinase-dependent activator of tyrosine and tryptophan hydroxylases. PNAS 85 (19): 7084–7087PubMedGoogle Scholar
  25. Ischimura T, Sugano H, Kuwano R, Sunaya T, Okuyama T, Isobe T (1991) Widespread distribution of the 14-3-3 protein in vertebrate brains and bovine tissues: correlation with the distribution of calcium-dependent protein kinases. J Neurochem 56: 1449–1451Google Scholar
  26. Ichimura-Ohshima Y, Mori K, Ichimura T, Araki K, Takahashi Y, Isobe T, Minoshima S, Fukuyama R, Shimizu N, Kuwano R (1992) cDNA cloning and chromosome assignment of the gene for human brain 14-3-3 protein eta-chain. J Neurosci Res 31: 600–605CrossRefPubMedGoogle Scholar
  27. Irie K, Gotoh Y, Yashar BM, Errede B, Nishida E, Matsumoto K (1994) Stimulatory effects of yeast and mammalian 14-3-3 proteins on the Raf protein kinase. Science 265: 1716–1719PubMedGoogle Scholar
  28. Isobe T, Ichimura T, Sunaya T, Okuyama T, Takahashi N, Kuwano R, Rosenfeld G, Takahashi Y (1991) Distinct forms of the protein kinase dependent activator of tyrosine and tryptophan hydroxylases. J Mol Biol 217: 125–132CrossRefPubMedGoogle Scholar
  29. Jarillo JA, Capel J, Leyva A, Martinez-Zapater JM, Salinas EBJ (1994) Two related low-temperature-inducible genes of arabidopsis encode proteins showing high homology to 14-3-3 proteins, a family of putative kinase regulators. Plant Mol Biol 25 (4): 693–704CrossRefPubMedGoogle Scholar
  30. Jones DH, Ley S, Aitken A (1995a) Isoforms of 14-3-3 protein can form homo- and heterodimers in vivo and in vitro: implications for function as adapter proteins. FEBS Lett 368: 55–58CrossRefPubMedGoogle Scholar
  31. Jones DH, Martin H, Madrazo J, Robinson KA, Neilsen P, Roseboom PH, Patel Y, Howell SA, Aitken A (1995b) Expression and structural analysis of 14-3-3 proteins. J Mol Biol 245: 375–384PubMedGoogle Scholar
  32. Korthout HA, deBoer AH (1994) A fusicoccin binding protein belongs to the family of 14-3-3 brain protein homologs. Plant Cell 6: 1681–1692.CrossRefPubMedGoogle Scholar
  33. Kidou S, Umeda M, Kato A, Uchimiya H (1993) Isolation and characterization of a rice cDNA similar to the bovine brain-specific 14-3-3 protein gene. Plant Mol Biol 21: 191–194CrossRefPubMedGoogle Scholar
  34. Laughner B, Lawrence SD, Ferl RJ (1994) Two tomato fruit homologs of 14-3-3 mammalian brain proteins. Plant Physiol 105 (4): 1457–1458CrossRefPubMedGoogle Scholar
  35. Leffers H, Madsen P, Rasmussen HH, Honore B, Andersen AH, Walbum E, Vandekerckhove J, Celis JE (1993) Molecular cloning and expression of the transformation sensitive epithelial with the G-box marker stratifin. A member of a protein family involved in the protein kinase C signalling pathway. J Mol Biol 231: 982–998CrossRefPubMedGoogle Scholar
  36. Liu D, Bienkowska J, Petosa C, Collier RJ, Fu H, Liddington R (1995) Crystal structure of the zeta isoform of the 14-3-3 protein. Nature 376: 191–194CrossRefPubMedGoogle Scholar
  37. Lu GH, Delisle AJ, deVetten NC, Ferl RJ (1992) Brain proteins in plants—an Arabidopsis homolog to neurotransmitter pathway activators is part of a DNA binding complex. PNAS 89 (23): 1490–1494Google Scholar
  38. Heusden GP, Ferl RJ (1994a) A single Arabidopsis GF14 isoform possesses biochemical characteristics of diverse 14-3-3 homologues. Plant Mol Biol 25 (4): 659–673PubMedGoogle Scholar
  39. Lu G, Sehnke PC, Ferl RJ (1994b) Phosphorylation and calcium binding properties of an Arabidopsis GF14 brain protein homolog. Plant Cell 6 (4): 501–510CrossRefPubMedGoogle Scholar
  40. Markiewicz E, Rzepecki R, Zopa S (1994) Molecular cloning and sequencing of the cDNA sequence of the cDNA encoding plant nuclear matrix endonuclease. Acta Biochim Pol 41: 137–138PubMedGoogle Scholar
  41. Martens GJM, Piosik PA, Danen EHJ (1992) Evolutionary conservation of the 14-3-3 protein. Biochim Biophys Res Commun 184: 1456–1459CrossRefGoogle Scholar
  42. Martin H, Patel Y, Jones D, Howell S, Robinson K, Aitken A (1993) Antibodies against the major brain isoforms of 14-3-3 protein: an antibody specitic for the N-acetylated amino-terminus of a protein. FEBS Lett 331: 296–303CrossRefPubMedGoogle Scholar
  43. McConnell JE, Armstrong JF, Hodges PE, Bard JB (1995) The mouse 14-3-3 epsilon isoform, a kinase regulator whose expression pattern is modulated in mesenchyme and neuronal differentiation. Dev Biol 169 (1): 218–228CrossRefPubMedGoogle Scholar
  44. Moore BW, Perez VJ (1967) Specific acidic proteins of the nervous system. In: Carlson FD (ed) Physiological and biochemical aspects of nervous integration. Prentice-Hall, Englewood Cliffs, NJ, pp 343–359Google Scholar
  45. Morgan A, Burgoyne RD (1992) Interaction between protein kinase C and Exo 1 (14-3-3 protein) and its relevance to exocytosis in permeabilized adrenal chromaffin cells. Biochem J 286: 807–811PubMedGoogle Scholar
  46. Morrison D (1994) 14-3-3 modulators of signaling proteins? Science 266: 56–57PubMedGoogle Scholar
  47. Nielsen PJ (1991) Primary structure of a human protein kinase regulator protein. Biochim Biophys Acta 1088: 425–428PubMedGoogle Scholar
  48. Oecking C, Eckerskorn C, Weiler EW (1994) The fusicoccin receptor of plants is a member of the 14-3-3 superfamily of eukaryotic regulatory proteins. FEBS Lett 352: 163–166CrossRefPubMedGoogle Scholar
  49. Pallas DC, Fu H, Haehnel LC, Weller W, Collier RJ, Roberts TM (1994) Association of polyomavirus middle tumor antigen with 14-3-3 proteins. Science 265: 535–537PubMedGoogle Scholar
  50. Prasad GL, Valverius EM, McDuffie E, Cooper HL (1992) Complementary DNA cloning of a novel epithelial cell marker protein, HME1, that may be down-regulated in neoplastic mammary cells. Cell Growth Differ 3 (8): 507–513PubMedGoogle Scholar
  51. Reuther GW, Fu H, Cripe LD, Collier RJ, Pendergast AM (1994) Association of the protein kinase c-Bcr and Bcr-Abl with proteins of the 14-3-3 family. Science 266: 129–133PubMedGoogle Scholar
  52. Roseboom PH, Weller JL, Babila T, Aitken A, Sellers LA, Moffett JR, Namboodiri MA, Klein DC (1994) Cloning and characterization of the epsilon and zeta isoforms of the 14-3-3 proteins. DNA Cell Biol 13 (6): 629–640PubMedGoogle Scholar
  53. Roth D, Morgan A, Martin H, Jones D, Martens GJ, Aitken A, Burgoyne RD (1994) Characterization of 14-3-3 proteins in adrenal chromaffin cells and demonstration of isoform-specific phospholipid binding. Biochem J 301: 305–310PubMedGoogle Scholar
  54. Stankovic B, Garic-Stankovic A, Smith CM, Davies E (1995) Isolation, sequencing, and analysis of a 14-3-3 brain protein homolog from pea (Pisum sativum L.). Plant Physiol 107 (4): 1481–1482CrossRefPubMedGoogle Scholar
  55. Swanson KD, Ganguly R (1992) Characterization of aDrosophila melanogaster gene similar to the mammalian genes encoding the tyrosine/tryptophan hydroxylase activator and protein kinase C inhibitor proteins. Gene 113: 183–190CrossRefPubMedGoogle Scholar
  56. Tanji M, Horwitz R, Rosenfeld G, Waymire JC (1994) Activation of protein kinase C by purified bovine brain 14-3-3: comparison with tyrosine hydroxylase activation. J Neurochem 63 (5): 1908–1916PubMedGoogle Scholar
  57. Toker A, Sellers LA, Amess B, Patel Y, Harris A, Aitken A (1992) Multiple isoforms of a protein kinase C inhibitor (KCIP/14-3-3) from sheep brain: amino acid sequence of phosphorylated forms. Eur J Biochem 206: 453–461CrossRefPubMedGoogle Scholar
  58. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choices. Nucleic Acids Res 22: 4673–4680PubMedGoogle Scholar
  59. van Heusden GPH, Wenzel TJ, Lagendijk EL, de Steensma HY, van den Berg JA (1992) Characterization of the yeast BMH1 gene encoding a putative protein homologous to mammalian protein kinase II activators and proteins kinase C inhibitors. FEBS Lett 302 (2): 145–150PubMedGoogle Scholar
  60. van Heusden GPH, Griffiths DJ, Ford JC, Chin A, Woeng TF, Schrader PA, Carr AM, Steensma HY (1995) The 14-3-3 proteins encoded by the BMH1 and BMH2 genes are essential in the yeastSaccharomyces cerevisiae and can be replaced by a plant homologue. Eur J Biochem 229 (1): 45–53CrossRefPubMedGoogle Scholar
  61. Wang W, Shakes DC (1994) Isolation and sequence analysis of aCaenorhabditis elegans cDNA which encodes a 14-3-3 homologue. Gene 147: 215–218CrossRefPubMedGoogle Scholar
  62. Watanabe M, Isobe T, Okuyama T, Ichimura T, Kuwano R, Takahashi Y, Kondo H (1991) Molecular cloning of cDNA to rat 14-3-3 eta chain polypeptide and the neuronal expression of the mRNA in the central nervous system. Mol Brain Res 10 (2): 151–158CrossRefPubMedGoogle Scholar
  63. Watanabe M, Isobe T, Ichimura T, Kuwano R, Takahashi Y, Kondo H (1993a) Molecular cloning of rat cDNAs for beta and gamma subtypes of 14-3-3 protein and developmental change in expression of their mRNAs in the nervous system. Mol Brain Res 17 (1–2): 135–146PubMedGoogle Scholar
  64. Watanabe M, Isobe T, Ichimura, Kuwano R, Takahashi Y, Kondo H (1993b) Developmental regulation of neuronal expression for the eta subtype of the 14-3-3 protein, a putative regulatory protein for protein kinase C. Dev Brain Res 73: 225–235CrossRefGoogle Scholar
  65. Watanabe M, Isobe T, Ichimura T, Kuwano R, Takahashi Y, Kondo H, Inoue Y (1994) Molecular cloning of rat cDNAs for the zeta and theta subtypes of 14-3-3 protein and differential distributions of their mRNAs in brain. Mol Brain Res 25 (1–2): 113–121PubMedGoogle Scholar
  66. Xiao B, Smerdon SJ, Johns DH, Dodson GG, Soneji Y, Aitken A, Gamblin SJ (1995) Structure of a 14-3-3 protein and implications for coordination of multiple signalling pathways. Nature 376: 188–191CrossRefPubMedGoogle Scholar
  67. Zupan LA, Steffens DL, Berry CA, Landt M, Gross RW (1992) Cloning and expression of a human 14-3-3 protein mediating phospholipolysis. J Biol Chem 13: 8707–8710Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1996

Authors and Affiliations

  • Wenfu Wang
    • 1
  • Diane C. Shakes
    • 2
  1. 1.Department of BiologyUniversity of HoustonHoustonUSA
  2. 2.Department of BiologyCollege of William and MaryWilliamsburgUSA

Personalised recommendations