Molecular cloning of aTrypanosoma cruzi cDNA encoding a protein homologous with mammalian elongation factor 1?

Plumas-Marty, Béatrice; Schöneck, Ralf; Billaut-Mulot, Odile; Taibi, A.; Càpron, André; Ma, Ouaissi

doi:10.1007/bf00933014

Cited by 10 publications

(8 citation statements)

References 14 publications

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“…The overall similarity (about 61%; fig 4) shown between TcEF-17and mammalian EF-17is of the same order as that observed between EF-lfl proteins of different species and a T cruzi protein homologous to mammalian elongation factor lfl (TcEF-lf125 and TcEF-lfl30) [19]. Moreover, three putative recognition sequences for p34 ede2 kinase were present in TcEF-1z respectively at positions 51 (CSPC), 90 (RTPL), and 265 (PSPF) (fig 1).…”

Section: Resultssupporting

confidence: 65%

“…Figure 5 shows a close c o r r e s p o n d e n c e of the arrangement of hydrophobic clusters and proline residues. Moreover, the hydrophilic region was clearly evidenced in the two sequences• In our original study, we have isolated TcGBP antigens using binding to glutathione matrix [19]. Comparison of TcEF-17 sequence with available nucleotide or amino acid sequences corresponding to proteins related to the glutathione cycle including GSTs, revealed some limited similarity (30%) between TcEF-17and fruit fly GST27 (data not shown).…”

Section: Resultsmentioning

confidence: 96%

“…2. In a previous study, we isolated TcGBP antigens using binding to glutathione matrix [19]. Comparison of TcEF-1fl25 and TcEF-lfl30 sequences with available nucleotide or amino acid sequences corresponding to proteins related to the glutathione cycle including GSTs did not reveal any significant homology.…”

Section: Notes Added In Proofmentioning

confidence: 95%

“…In the mammalian host, the parasite cycles between intracellular reproductive amastigotes and the released infective trypomastigote forms. Apart from the preliminary observation of TcEF-1)' [2] and TcEF-lfl [19], no evidence documenting the existence of either EF-lfl or )'in Kinetoplastidae has yet been reported. In this paper we present detailed analysis of T cruzi cDNA encoding TcEF-1 y homologue and show that antibodies to the fusion peptide reacted against macrophage cell line J774 extracts, suggesting therefore that the EF-1)'is highly conserved across evolution.…”

Section: Introductionmentioning

confidence: 91%

“…Rat antibodies against a complex of three parasite proteins with molecular masses of 25, 30 and 45 kDa named TcGBP and rabbit antibodies to a mixture of 25 and 30 kDa proteins (TcEF-I fl 25-30), produced in pGEX-2T vector as fusion peptides with Schistosoma japonicum glutathione S-transferase (Sj26), were obtained as described [18,19]. After the double immunoscreening strategy (see below), pBluescript inserts coding for the 45 kDa protein (TcEF-17) were subcloned into EcoRI, XhoI restriction sites of pGEX-4T-3 (Pharmacia, Uppsala, Sweden) vector.…”

Section: Antibodiesmentioning

confidence: 99%

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Molecular and immunological characterization of a Trypanosoma cruzi protein homologous to mammalian elongation factor 1 gamma

Billaut-Mulot

Schöneck

Fernández-Gómez

et al. 1994

Biology of the Cell

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In previous studies, we reported the characterization of three Trypanosoma cruzi proteins with molecular masses of 45, 30 and 25 kDa eluted from a glutathione agarose column (these proteins were named TcGBP). Using antibodies against TcGBP native proteins we could isolate from a lambda ZAPII epimastigote cDNA library cDNA clones encoding the 30 and 25 kDa proteins. Comparison of the two sequences with amino acid sequences in several data banks revealed that both protein sequences were highly homologous to human and Artemia salina elongation factor 1 beta. Thus, the proteins were named TcEF-1 beta 25 and TcEF-1 beta 30. In the present study we used a double immunoscreening strategy that allowed us to isolate a cDNA clone corresponding to the 45 kDa protein. The protein sequence revealed 31% identity and 61% homology with human and Artemia salina EF1 gamma and therefore was named TcEF-1 gamma. Moreover, three putative phosphorylation sites at position 51 (CSPC), at position 90 (RTPL) and at position 265 (PSPF) were found in the TcEF-1 gamma sequence. These sites are compatible with the notion that TcEF-1 gamma could be the target of phosphorylation by protein kinase(s). Random primed cDNA hybridized with a single 1.4 kb mRNA found in epimastigote, trypomastigote and amastigote forms. In addition, Southern blot analysis of genomic DNA suggested that the protein is encoded by a single gene. The TcEF-1 gamma cDNA was subcloned into the pGEX-4T-3 vector for expression in Escherichia coli.(ABSTRACT TRUNCATED AT 250 WORDS)

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