Abstract
The techniques of homology cloning and anchored PCR were used to clone the elongation factor 2 (EF-2) gene from black tiger shrimp (Penaeus monodon). The full length cDNA of black tiger shrimp EF-2 (btsEF-2) contained a 5′ untranslated region (UTR) of 73 bp, an ORF of 2541 bp encoding a polypeptide of 846 amino acids with an estimated molecular mass of 95 kDa, and a 3( UTR of 112 bp. The searches for protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of btsEF-2 was homological to the EF-2 of other species and even the mammalians. The conserved signature sequence of EF-2 gene family, GTPase effector domain and ADP-ribosylation domain were found in the btsEF-2 deduced amino acid sequence. The temporal expressions of gene in the different ovarian stages were measured by real time PCR. The mRNA expressions of the gene were constitutively expressed in ovary and different during the maturation stages. The result indicated that EF-2 gene was constitutively expressed and could play a critical role in the ovarian maturation stage.
Similar content being viewed by others
References
Mathews MB, Sonenberg N, Hershey JWB (2000) Origins and principles of translational control. In: Sonenberg N, Hershey JWB, Mathews MB (eds) Translational Control of Gene Expression. Cold Spring Harbor Laboratory Press, New York, pp 1–32
Tania MM, James DF (2004) Identification of a developmentally regulated translation elongation factor 2 in Tetrahymena thermophila. Gene 326:97–105
Weissbach H, Ochoa S (1976) Soluble factors required for eukaryotic protein synthesis. Annu Rev Biochem 45:191–216
Lim EJ, Kim, ChW (2007) Functional characterization of the promoter region of the chicken elongation factor-2 gene. Gene 386:183–190
Nairn AC, Palfrey HC (1987) Identification of the major Mr 1,000,000 substrate for calmodulin dependent protein kinase III in mammalian cells as elongation factor 2. J Biol Chem 262:17299–17303
Redpath NC, Price NT, Severinov KV, Proud CG (1993) Regulation of elongation factor-2 by multisite phosphorylation. Eur J Biochem 213:689–699
Carlberg U, Nilsson A, Nygard O (1990) Functional properties of phosphorylated elongation factor 2. Eur J Biochem 191:639–645
Ortiz PA, Kinzy TG (2005) Dominant-negative mutant phenotypes and the regulation of translation elongation factor 2 levels in yeast. Nucleic Acids Res 33:5740–5748
Redpath NC, Foulstone EJ, Proud CG (1996) Regulation of translation elongation factor-2 by insulin via a rapamycin sensitive pathway. EMBO J 15:2291–2297
Campbell LE, Wang X, Proud CG (1999) Nutrients differentially regulate multiple translation factors and their control by insulin. Biochem J 344:433–441
Fendrick JL, Iglewski WJ, Moehring JM, Moehring TJ (1992) Characterization of the endogeneous ADP-ribosylation of mutant and wild-type elongation factor 2 in eukaryotic cells. Eur J Biochem 205:25–31
Browne GJ, Proud CG (2002) Regulation of peptide-chain elongation in mammalian cells. Eur J Biochem 269:5360–5368
Sans MD, Xie Q, Williams JA (2004) Regulation of translation elongation and phosphorylation of eEF2 in rat pancreatic acini. Biochem Biophys Res Commun 319:144–151 (Jun.)
Oleinikov AV, Jokhadze GG, Alakhov, Yu B (1989) Primary structure of rat liver elongation factor 2 deduced from the cDNA sequence. FEBS Lett 248:131–136
Kohno K, Uchida T, Ohkubot H, Nakanishid S, Nakanishi T, Fukui T, Ohtsuka E, Ikehara M, Okadat Y (1986) Amino acid sequence of mammalian elongation factor 2 deduced from the cDNA sequence: Homology with GTP-binding proteins. Proc Natl Acad Sci USA 83:4978–4982
Rapp G, Klaudiny J, Hagendorff G, Luck MR, Scheit KH (1989) Complete sequence of the coding region of human elongation factor 2 (EF-2) by enzymatic amplification of cDNA from human ovarian granulosa cells. Biol Chem Hoppe Seyler 370:1071–1075
Kapaettu Satyamoorthy, Chin CHowe (1997) The mouse elongation factor-2 gene: isolation and characterization of the promoter. DNA Cell Biol 16:402–412
Kim CW, Jung EJ, Kim YW, Kang KR (1993) Molecular cloning of chicken elongation factor 2 (EF-2): sequence comparison with mammalian EF-2 and its expression in the early development stages of the embryos. Mol Cells 3:27–33
Zhang SH, Yao JH, Song HD, Wang L, Xue JL (2006) Cloning, expression and functional study of translation elongation factor 2 (EF-2) in zebrafish Int. J Dev Biol 50:399–403
Huang JH, Zhou FL, Ma ZM, Jiang SG (2005) Morphological and histological observation on ovary development of Penaeus monodon from northern South China Sea. J Trop Oceanogr 25(3):47–52
Saitou N, Nei M (1987) The neighbour-joining method-a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
la Cour TF, Nyborg J, Thirup S, Clark BF (1985) Structural details of the binding of guanosine diphosphate to elongation factor Tu from E. coli as studied by X-ray crystallography. EMBO J 4:2385–2388
Kohno K, Uchida T (1987) Highly frequent single amino acid substitution in mammalian elongation factor 2 (EF-2) results in expression of resistance to EF-2-ADP-ribosylating toxins. J Biol Chem 262:12298–12305
Kimata Y, Harashima S, Kohno K (1993) Expression of non-ADP-ribosylatable, diphtheria toxin-resistant elongation factor 2 in Saccharomyces cerevisiae. Biochem Biophys Res Commun 191:1145–1151
Wang X, Campbell L, Miller C, Proud C (1998) Amino acid availability regulates p70 S6 kinase and multiple translation factors. Biochem J 334:261–267
Perentesis JP, Phan LD, Gleason WB, LaPorte DC, Livingston DM, Bodley JW (1992) Saccharomyces cerevisiae elongation factor 2. J Biol Chem 267:1190–1197
Lasko P (2000) The Drosophila melanogaster genome: translation factors and RNA binding proteins. J Cell Biol 150:51–56
Hovemman B, Richter S, Waldorf U, Cziepluch C (1988) Two genes encode related cytoplasmic elongation factors 1a (EF-1a) in Drosophila melanogaster with continuous and stage specific expression. Nucleic Acids Res 16:3175–3194
Dje´ MK, Mazabraud A, Viel A, le Marie M, Denis H, Crawford E, Brown DD (1990) Three genes under different developmental control encode elongation factor 1-a in Xenopus laevis. Nucleic Acids Res 18:3489–3493
Acknowledgments
This study was supported by National nature foundation of China (No.30571447), National “863” Project of China (No. 2003 AA603120 ) and Agriculture Department Project of China(06–05-01B).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Qiu, L., Jiang, S., Zhou, F. et al. Molecular cloning of the black tiger shrimp (Penaeus monodon) elongation factor 2 (EF-2): sequence analysis and its expression on the ovarian maturation stage. Mol Biol Rep 35, 431–438 (2008). https://doi.org/10.1007/s11033-007-9103-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-007-9103-5