1887

Abstract

This paper reports the identification of Ehp53, a p53-like 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 nuclear extracts and oli-p53. Additionally, 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 and . 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 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 Zn-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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.25892-0
2003-04-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/4/mic149885.html?itemId=/content/journal/micro/10.1099/mic.0.25892-0&mimeType=html&fmt=ahah

References

  1. Ankri S., Stolarsky T., Mirelman D. 1998; Antisense inhibition of expression of cysteine proteinases does not affect Entamoeba histolytica cytopathic or haemolytic activity but inhibits phagocytosis. Mol Microbiol 28:777–785
    [Google Scholar]
  2. Ankri S., Stolarsky T., Bracha R., Padilla-Vaca F., Mirelman D. 1999; Antisense inhibition of expression of cysteine proteinases affects Entamoeba histolytica -induced formation of liver abscess in hamsters. Infect Immun 67:421–422
    [Google Scholar]
  3. Banks L., Matlashewsky G., Crawford L. 1986; Isolation of human-p53-specific monoclonal antibodies and their use in studies of human p53 expression. Eur J Biochem 159:529–534
    [Google Scholar]
  4. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  5. Brodsky M. H., Nordstrom W., Tsang G., Kwan E., Rubin G. M., Abrams J. M. 2000; Drosophila p53 binds a damage response element at the reaper locus. Cell 101:103–113
    [Google Scholar]
  6. Clark C. G., Diamond L. S. 1991; The Laredo strain and other “ Entamoeba histolytica -like” amoebae are E. moshkovskii . Mol Biochem Parasitol 46:11–18
    [Google Scholar]
  7. Cox L. S., Lane D. P. 1995; Tumour suppressors, kinases and clamps: how p53 regulates the cell cycle in response to DNA damage. BioAssays 17:501–508
    [Google Scholar]
  8. Cox L. S., Midgley C. A., Lane D. P. 1994; Xenopus p53 is biochemically similar to the human tumor suppressor protein p53 and is induced upon DNA damage in somatic cells. Oncogene 9:2951–2959
    [Google Scholar]
  9. Descoteaux S., Ayala P., Samuelson J., Orozco E. 1992; Primary sequences of two P-glycoprotein genes of Entamoeba histolytica . Mol Biochem Parasitol 54:200–202
    [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
    [Google Scholar]
  11. Diamond L. S., Harlow D. R., Cunick C. C. 1978; A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba . Trans R Soc Trop Med Hyg 72:431–432
    [Google Scholar]
  12. El-Deiry W. S., Kern S. E., Pietenpol J. A., Kinzler K. W., Vogelstein B. 1992; Definition of a consensus binding site for p53. Nat Genet 1:45–49
    [Google Scholar]
  13. Enoch T., Norbury C. 1995; Cellular responses to DNA damage: cell-cycle checkpoints, apoptosis and the roles of p53 and ATM. Trends Biochem Sci 20:426–430
    [Google Scholar]
  14. García-Rivera G., Gómez C., Pérez D. G., Flores E., Rodríguez M. A., Orozco E. 2000; Structural characterization of the 5′-flanking regions of the Entamoeba histolytica Ehcp112 and Ehadh112 genes. Arch Med Res 31:S303–S304
    [Google Scholar]
  15. Gómez C., Pérez D. G., Lopez-Bayghen E., Orozco E. 1998; Transcriptional analysis of the EhPgp1 promoter of Entamoeba histolytica multidrug-resistant mutant. J Biol Chem 273:7277–7284
    [Google Scholar]
  16. Guimaraes S., Urdaneta H., Silva E. F., Tavares C. A. 1991; Entamoeba histolytica : antigenic characterization of axenic strains from Brazil. Rev Inst Med Trop Sao Paulo 33:6–11
    [Google Scholar]
  17. Kumaravel S., Wafik E. S. 2000; Tumor suppressor p53: regulation and function. Front Biosci 5:424–437
    [Google Scholar]
  18. Loidl A., Loidl P. 1996; Oncogene and tumor suppressor gene-related proteins in plants and fungi. Crit Rev Oncogenesis 7:49–64
    [Google Scholar]
  19. Lu W., Pochampally R., Chen L., Traidej M., Wang Y., Chen J. 2000; Nuclear exclusion of p53 in a subset of tumors requires MDM2 function. Oncogene 19:232–240
    [Google Scholar]
  20. Luna-Arias J. P., Hernandez-Rivas R., De Dios-Bravo G., García J., Mendoza L., Orozco E. 1999; The TATA-box binding protein of Entamoeba histolytica : cloning of the gene and location of the protein by immunofluorescence and confocal microscopy. Microbiology 145:33–40
    [Google Scholar]
  21. Maltzman W., Czyzyk L. 1984; UV irradiation stimulates levels of p53 cellular tumor antigen in nontransformed mouse cells. Mol Cell Biol 4:1689–1694
    [Google Scholar]
  22. Marchat L. A., Gómez C., Pérez D. G., Paz F., Mendoza L., Orozco E. 2002; The CCAAT/enhancer binding protein sites are cis -activator elements of the Entamoeba histolytica Ehpgp1 ( mdr -like) gene expresion. Cell Microbiol 4:725–737
    [Google Scholar]
  23. Middeler G., Zerf K., Jenovai S., Thulig A., Tschodrich-Rotter M., Kubitscheck U., Peters R. 1997; The tumor suppressor p53 is subject to both nuclear import and export, and both are fast, energy-dependent and lectin-inhibited. Oncogene 14:1407–1417
    [Google Scholar]
  24. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. 1977; High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142
    [Google Scholar]
  25. Ollman M., Young L. M., Di Como C. J. 10 other authors 2000; Drosophila p53 is a structural and functional homolog of the tumor suppressor p53. Cell 101:91–101
    [Google Scholar]
  26. Orozco E., Guarneros G., Martínez-Palomo A., Sánchez T. 1983; Entamoeba histolytica . Phagocytosis as a virulence factor. J Exp Med 158:1511–1521
    [Google Scholar]
  27. Orozco E., Gharaibeh R., Riverón A. M., Delgadillo D. M., Mercado M., Sanchez T. 1997; A novel cytoplasmic structure containing DNA networks in Entamoeba histolytica trophozoites. Mol Gen Genet 254:250–257
    [Google Scholar]
  28. Pearson W. R., Lipman D. J. 1988; Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A 85:2444–2448
    [Google Scholar]
  29. Pérez D. G., Gómez C., Lopez-Bayghen E., Tannich E., Orozco E. 1998; Transcriptional analysis of the EhPgp5 promoter of the Entamoeba histolytica multidrug-resistant mutant. J Biol Chem 273:7285–7292
    [Google Scholar]
  30. Petri W. A., Smith R. D., Schlesinger P. H., Ravdin J. I. 1987; Isolation of the galactose binding lectin which mediates the in vitro adherence of Entamoeba histolytica . J Clin Invest 80:1238–1244
    [Google Scholar]
  31. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  32. Sánchez M. A., Peattie D. A., Wirth D., Orozco E. 1994; Cloning, genomic organization and transcription of the Entamoeba histolytica α -tubulin encoding gene. Gene 146:239–244
    [Google Scholar]
  33. Schaenman J. M., Gilchrist C. A., Mann B. J., Petri W. A. Jr 2001; Identification of two E. histolytica sequence-specific URE4 enhancer binding proteins with homology to the RNA-binding motif RRM. J Biol Chem 276:1602–1609
    [Google Scholar]
  34. Scheffner M., Werness B. A., Huibregtse J. M., Levine A. J., Howley P. M. 1990; The E6 oncoprotein encoded by human papillomavirus type 16 and 18 promotes the degradation of p53. Cell 63:1129–1136
    [Google Scholar]
  35. Schmale H., Bamberger C. 1997; A novel protein with strong homology to the tumor suppressor p53. Oncogene 15:1363–1367
    [Google Scholar]
  36. Shaulsky G., Ben-Ze'ev A., Rotter V. 1990a; Subcellular distribution of the p53 protein during the cell cycle of Balb/c 3T3 cells. Oncogene 6:1707–1711
    [Google Scholar]
  37. Shaulsky G., Goldfinger N., Ben-Ze'ev A., Rotter V. 1990b; Nuclear accumulation of p53 protein is mediated by several nuclear localization signals and plays a role in tumorigenesis. Mol Cell Biol 10:6565–6577
    [Google Scholar]
  38. Smith M. L., Chen I. T., Zhan Q., Bae I., Chen C. Y., Gilmer T. M., Kastan M. B., O'Connor P. M., Fomace A. J. Jr 1994; Interaction of the p53-regulated protein GADD45 with proliferating cell nuclear antigen. Science 266:1376–1380
    [Google Scholar]
  39. Stommel J. M., Marchenko N. D., Jimenez G. S., Moll U. M., Hope T. J., Wahl G. M. 1999; A leucine-rich nuclear export signal in the p53 tetramerization domain: regulation of subcellular localization and p53 activity by NES masking. EMBO J 18:1660–1672
    [Google Scholar]
  40. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4356
    [Google Scholar]
  41. Van Beneden R. J., Walker C. W., Laughner E. S. 1997; Characterization of gene expression of a p53 homologue in the soft-shell clam ( Mya arenaria ). Mol Mar Biol Biotechnol 6:116–122
    [Google Scholar]
  42. WHO 1997; Amoebiasis. WHO Weekly Epidemiol Record 72:97–100
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.25892-0
Loading
/content/journal/micro/10.1099/mic.0.25892-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error