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Microbiology 149 (2003), 885-893; DOI  10.1099/mic.0.25892-0
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Microbiology 149 (2003), 885-893; DOI  10.1099/mic.0.25892-0
© 2003 Society for General Microbiology

Ehp53, an Entamoeba histolytica protein, ancestor of the mammalian tumour suppressor p53

L. Mendoza1, E. Orozco1, M. A. Rodríguez1, G. García-Rivera1, T. Sánchez1, E. García2 and P. Gariglio2

1 Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), AP 14-740, DF 07000, Mexico
2 Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), AP 14-740, DF 07000, Mexico

Correspondence
E. Orozco
esther{at}mail.cinvestav.mx

This paper reports the identification of Ehp53, a p53-like Entamoeba histolytica protein, which binds to the human p53 DNA consensus sequence (oli-p53). Monoclonal antibodies against p53 (Ab-1 and Ab-2) recognized a single 53 kDa spot in two-dimensional gels and inhibited the formation of complexes produced by E. histolytica nuclear extracts and oli-p53. Additionally, E. histolytica gene promoter sequences with high homology to oli-p53 formed complexes with nuclear proteins that were abolished by oli-p53. Ehp53 protein levels increased in UV-irradiated trophozoites. This protein was also detected in Entamoeba moshkovskii and Entamoeba invadens. By confocal microscopy, Ehp53 was located in the nuclei, EhkO organelles and cytoplasm. The Ehp53-encoding gene was cloned and its predicted amino acid sequence showed 30–54 % and 50–57 % homology with important domains of the human and the Drosophila melanogaster p53 proteins, respectively. This homology included the tetramerization domain, the nuclear export signal and a nuclear localization signal. Ehp53 also contains seven of the eight DNA-binding residues and two of the four Zn2+-binding sites described for p53. A recombinant Ehp53 was recognized by Ab-2. Ehp53 is believed to be the first p53-like protein found in protozoa and may be the evolutionary ancestor of the mammalian p53.

Abbreviations: Ab-1 and Ab-2, monoclonal antibodies against the human p53; CE, cytoplasmic extracts; EhkO, E. histolytica kinetoplast-like organelle; NE, nuclear extracts; oli-p53, DNA consensus sequence for mammalian p53; oli-Eh112, oligonucleotide from E. histolytica with homologous sequence to oli-p53

The GenBank accession number for the sequence reported in this paper is AJ489250.




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