Abstract
Previous studies have demonstrated that 14-3-3 proteins exist in all the eukaryotic organisms studied; however, studies on the 14-3-3 proteins have not been involved in the halotolerant, unicellular green alga Dunaliella salina so far. In the present study, a cDNA encoding 14-3-3 protein of D. salina was cloned and sequenced by PCR and rapid amplification of cDNA end (RACE) technique based on homologous sequences of the 14-3-3 proteins found in other organisms. The cloned cDNA of 1485 bp in length had a 29.2 kDa of molecular weight and contained a 774 bp of open reading frame encoding a polypeptide of 258 amino acids. Like the other 14-3-3 proteins, the deduced amino acid sequences of the D. salina 14-3-3 protein also contained two putative phosphorylation sites within the N-terminal region (positions 62 and 67). Furthermore, an EF hand motif characteristic for Ca2+-binding sites was located within the C-terminal part of this polypeptide (positions 208–219). Analysis of bioinformatics revealed that the 14-3-3 protein of D. salina shared homology with that of other organisms. Real-time quantitative PCR demonstrated that expression of the 14-3-3 protein gene is cell cycle-dependent.
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Abbreviations
- ORF:
-
Open reading frame
- RACE:
-
Rapid amplification of cDNA ends
- RT-PCR:
-
Reverse transcriptase-polymerase chain reaction
References
Liebich I, Voigt J (1995) A Chlamydomonas homologue to the 14-3-3 proteins: cDNA and deduced amino acid sequence. Biochim Biophys Acta 1263:79–85
Voigt J, Frank R (2003) 14-3-3 proteins are constituents of the insoluble glycoprotein framework of the chlamydomonas cell wall. Plant Cell 15:1399–1413
Moore BW, Perez VJ (1967) Specific acid proteins in the nervous system. In: Carlson FD (ed) Physiological and biochemical aspects of nervous integration. Prentice-Hall, Englewood Cliffs Prentice Hall, New Jersey, pp 343–359
Aitken A, Colling DB, Van Heusden BPH, Isobe T, Roseboom PH, Rosenfeld G, Soll J (1992) 14–3-3 proteins: a highly conserved, widespread family of eukaryotic proteins. Trends Biochem Sci 17:498–501
Ichimura 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 distributions of calcium-dependent protein kinases. J Neurochem 56:1449–1451
Tang SJ, Suen TC, McInnes RR, Buchwald M (1998) Association of the TLX-2 homeodomain and 14-3-3 eta signaling proteins. J Biol Chem 273:25356–25363
Morrison D (1994) 14-3-3: modulators of signaling protein. Science 266:56–57
Wilker EW, van Vugt MA, Artim SA, Huang PH, Petersen CP, Reinhardt HC, Feng Y, Sharp PA, Sonenberg N, White FM, Yaffe MB (2007) 14-3-3 sigma controls mitotic translation to facilitate cytokinesis. Nature 446:329–332
Yang X, Lee WH, Sobott F, Papagrigoriou E, Robinson CV, Grossmann JG, Sundstrom M, Doyle DA, Elkins JM (2006) Structural basis for protein-protein interactions in the 14-3-3 protein family. Proc Natl Acad Sci USA 103:17237–17242
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. J Biol Chem 270:5706–5709
Piotrowski M, Oecking C (1998) Five new 14-3-3 isoforms from Nicotiana tabacum L.: implications for the phylogeny of plant 14-3-3 proteins. Planta 204:127–130
Wang W, Shakes DC (1996) Molecular evolution of the 14-3-3 protein family. J Mol Evol 43:384–398
Vasara T, Keranen S, Penttila M, Saloheimo M (2002) Characterization of two 14-3-3 genes from Trichoderma reesei: interactions with yeast secretory pathway components. Biochim Biophys Acta 1590:27–40
Aitken A (2006) 14-3-3 proteins: a historic overview. Semin Cancer Biol 16:162–172
Wu K, Rooney MF, Ferl RJ (1997) The Arabidopsis 14-3-3 multigene family. Plant Physiol 114:1421–1431
Daugherty CJ, Ronney MF, Miller PW, Ferl RJ (1996) Molecular organization and tissue-specific expression of an Arabidopsis 14-3-3 gene. Plant Cell 8:1239–1248
Voigt J, Stevanovic S, Schirle M, Fausel M, Maier J, Adam KH, Marquardt O (2004) A 14-3-3 protein of Chlamydomonas reinhardtii associated with the endoplasmic reticulum: nucleotide sequence of the cDNA and the corresponding gene and derived amino acid sequence. Biochim Biophys Acta 1679:180–194
Voigt J, Liebich I, Kiess M, Frank R (2001) Subcellular distribution of 14-3-3 proteins in the unicellular green alga Chlamydomonas reinhardtii. Eur J Biochem 268:6449–6457
Wang T, Xue L, Hou W, Yang B, Cha Y, Ji X, Wang Y (2007) Increased expression of transgene in stably transformed cells of Dunaliella salina by matrix attachment regions. Appl Microbiol Biotechnol 76(3):651–657
Silflow CD, Youngblom J (1986) Chlamydomonas reinhardtii tubulin gene structure. Ann NY Acad Sci 466:18–30
Combet C, Blanchet C, Geourjon C, Deleage G (2000) Network protein sequence analysis. Trends Biochem Sci 25:147–150
Voigt J, Liebich I, Wöstemeyer J, Adam KH, Marquardt O (2000) Nucleotide sequence, genomic organization and cell-cycle-dependent expression of a Chlamydomonas 14-3-3 gene. Biochim Biophys Acta 1492(2–3):395–405
Acknowledgements
This work was supported by the grants from National Natural Science Foundation of China (No.30470030), and carried out in Henan Key Laboratory for Molecular Medicine.
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Wang, T., Xue, L., Ji, X. et al. Cloning and characterization of the 14-3-3 protein gene from the halotolerant alga Dunaliella salina . Mol Biol Rep 36, 207–214 (2009). https://doi.org/10.1007/s11033-007-9168-1
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DOI: https://doi.org/10.1007/s11033-007-9168-1