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Molecular Biology Reports

, Volume 39, Issue 4, pp 4545–4551 | Cite as

Transferrin regulates mRNA levels of a gene involved in iron utilization in Entamoeba histolytica

  • Cristhian Sánchez-Cruz
  • Mavil López-Casamichana
  • Areli Cruz-Castañeda
  • José de Jesús Olivares-Trejo
Article

Abstract

Entamoeba histolytica is a human pathogen, which can survive using haemoglobin (Hb) as only iron supply. Two probable haemophores (Ehhmbp26 and Ehhmbp45) are involved in iron acquisition in this parasite. However, mechanisms related to their transcriptional regulation have not been studied yet. In the present work, transcriptional profiles of both genes were evaluated in trophozoites cultures grown with different iron sources. ehhmbp26 gene was repressed totally by free iron, whereas ehhmbp45 gene showed clearly detectable mRNA levels. Expression profiles for both genes were significantly increased under iron privation condition. Interestingly, ehhmbp26 transcript was highly expressed by Holo-transferrin presence. This induction appears to be independent of direct contact between these proteins, because, in vitro assays evidenced that Ehhmbp26 protein was unable to bind this metalloprotein. Besides, in silico analysis of promoter nucleotide sequences of ehhmbp26 and ehhmbp45 genes revealed some distinctive core promoter elements described in E. histolytica and T-rich regions. Taking altogether these data suggest in E. histolytica dissimilar transcriptional mechanisms involved on iron acquisition control the expression of these genes, and they are unlike to those previously described for instance: in bacteria. Our findings evidenced this pathogen regulates the expression of ehhmbp26 and ehhmbp45 genes depending on the available iron supply, always ensuring the success of its infective process.

Keywords

Holo-Transferrin Transcription Iron-regulation Entamoeba histolytica Expression profile 

Notes

Acknowledgments

This work was supported by Consejo Nacional de Ciencia y Tecnología (CONACyT), México and Instituto de Ciencia y Tecnología del Distrito Federal (ICyT-DF).

References

  1. 1.
    Marquis JF, Gros P (2007) Intracellular Leishmania: your iron or mine? Trends Microbiol 15:93–95PubMedCrossRefGoogle Scholar
  2. 2.
    Perkins-Balding D, Ratliff-Griffin M, Stojiljkovic I (2004) Iron transport systems in Neisseria meningitidis. Microbiol Mol Biol Rev 68:154–171PubMedCrossRefGoogle Scholar
  3. 3.
    Wilson ME, Britigan BE (1998) Iron acquisition by parasitic protozoa. Parasitol Today 14:348–353PubMedCrossRefGoogle Scholar
  4. 4.
    Steverding D (2000) The transferrin receptor of Trypanosoma brucei. Parasitol Int 48:191–198PubMedCrossRefGoogle Scholar
  5. 5.
    Tsai CD, Liu HW, Tai JH (2002) Characterization of an iron-responsive promoter in the protozoan pathogen Trichomonas vaginalis. J Biol Chem 277:5153–5162PubMedCrossRefGoogle Scholar
  6. 6.
    Dey I, Keller K, Belley A, Chadee K (2003) Identification and characterization of a cyclooxygenase-like enzyme from Entamoeba histolytica. Proc Natl Acad Sci USA 100:13561–13566PubMedCrossRefGoogle Scholar
  7. 7.
    Reyes-Lopez M, Serrano-Luna JJ, Negrete-Abascal E, Leon-Sicairos N, Guerrero-Barrera AL, De la GM (2001) Entamoeba histolytica: transferrin binding proteins. Exp Parasitol 99:132–140PubMedCrossRefGoogle Scholar
  8. 8.
    Cruz-Castaneda A, Olivares-Trejo JJ (2008) Ehhmbp45 is a novel hemoglobin-binding protein identified in Entamoeba histolytica. FEBS Lett 582:2806–2810PubMedCrossRefGoogle Scholar
  9. 9.
    Cruz-Castaneda A, Hernandez-Sanchez J, Olivares-Trejo JJ (2009) Cloning and identification of a gene coding for a 26-kDa hemoglobin-binding protein from Entamoeba histolytica. Biochimie 91:383–389PubMedCrossRefGoogle Scholar
  10. 10.
    Cruz-Castaneda A, Lopez-Casamichana M, Olivares-Trejo JJ (2011) Entamoeba histolytica secretes two haem-binding proteins to scavenge haem. Biochem J 434:105–111PubMedCrossRefGoogle Scholar
  11. 11.
    Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72:431–432PubMedCrossRefGoogle Scholar
  12. 12.
    Rossi MS, Fetherston JD, Letoffe S, Carniel E, Perry RD, Ghigo JM (2001) Identification and characterization of the hemophore-dependent heme acquisition system of Yersinia pestis. Infect Immun 69:6707–6717PubMedCrossRefGoogle Scholar
  13. 13.
    Leon-Sicairos N, Reyes-Lopez M, Canizalez-Roman A, Bermudez-Cruz RM, Serrano-Luna J, Arroyo R, De la GM (2005) Human hololactoferrin: endocytosis and use as an iron source by the parasite Entamoeba histolytica. Microbiology 151:3859–3871PubMedCrossRefGoogle Scholar
  14. 14.
    Lopez-Soto F, Leon-Sicairos N, Reyes-Lopez M, Serrano-Luna J, Ordaz-Pichardo C, Pina-Vazquez C, Ortiz-Estrada G, De la GM (2009) Use and endocytosis of iron-containing proteins by Entamoeba histolytica trophozoites. Infect Genet Evol 9:1038–1050PubMedCrossRefGoogle Scholar
  15. 15.
    Flores-Soto E, zuara-Liceaga E, Lopez-Camarillo C, Orozco E (2005) The Entamoeba histolytica Ehcp112 gene has a distal and weak promoter. Exp Parasitol 110:286–291PubMedCrossRefGoogle Scholar
  16. 16.
    Hackney JA, Ehrenkaufer GM, Singh U (2007) Identification of putative transcriptional regulatory networks in Entamoeba histolytica using Bayesian inference. Nucleic Acids Res 35:2141–2152PubMedCrossRefGoogle Scholar
  17. 17.
    Perez DG, Gomez C, Lopez-Bayghen E, Tannich E, Orozco E (1998) Transcriptional analysis of the EhPgp5 promoter of Entamoeba histolytica multidrug-resistant mutant. J Biol Chem 273:7285–7292PubMedCrossRefGoogle Scholar
  18. 18.
    Purdy JE, Pho LT, Mann BJ, Petri WA Jr (1996) Upstream regulatory elements controlling expression of the Entamoeba histolytica lectin. Mol Biochem Parasitol 78:91–103PubMedCrossRefGoogle Scholar
  19. 19.
    Smith JM, Meerovitch E (1982) Specificity of iron requirements of Entamoeba histolytica in vitro. Arch Invest Med (Mex) 13(3):63–69Google Scholar
  20. 20.
    Buss H, Lioutas C, Dobinsky S, Nickel R, Tannich E (1995) Analysis of the 170-kDa lectin gene promoter of Entamoeba histolytica. Mol Biochem Parasitol 72:1–10PubMedCrossRefGoogle Scholar
  21. 21.
    Singh U, Rogers JB, Mann BJ, Petri WA Jr (1997) Transcription initiation is controlled by three core promoter elements in the hgl5 gene of the protozoan parasite Entamoeba histolytica. Proc Natl Acad Sci USA 94:8812–8817PubMedCrossRefGoogle Scholar
  22. 22.
    Romero-Diaz M, Gomez C, Lopez-Reyes I, Martinez MB, Orozco E, Rodriguez MA (2007) Structural and functional analysis of the Entamoeba histolytica EhrabB gene promoter. BMC Mol Biol 8:82PubMedCrossRefGoogle Scholar
  23. 23.
    Gomez C, Perez DG, Lopez-Bayghen E, Orozco E (1998) Transcriptional analysis of the EhPgp1 promoter of Entamoeba histolytica multidrug-resistant mutant. J Biol Chem 273:7277–7284PubMedCrossRefGoogle Scholar
  24. 24.
    Singh U, Rogers JB (1998) The novel core promoter element GAAC in the hgl5 gene of Entamoeba histolytica is able to direct a transcription start site independent of TATA or initiator regions. J Biol Chem 273:21663–21668PubMedCrossRefGoogle Scholar
  25. 25.
    Singh U, Gilchrist CA, Schaenman JM, Rogers JB, Hockensmith JW, Mann BJ, Petri WA (2002) Context-dependent roles of the Entamoeba histolytica core promoter element GAAC in transcriptional activation and protein complex assembly. Mol Biochem Parasitol 120:107–116PubMedCrossRefGoogle Scholar
  26. 26.
    Iyer LM, Anantharaman V, Wolf MY, Aravind L (2008) Comparative genomics of transcription factors and chromatin proteins in parasitic protists and other eukaryotes. Int J Parasitol 38:1–31PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Cristhian Sánchez-Cruz
    • 1
  • Mavil López-Casamichana
    • 1
  • Areli Cruz-Castañeda
    • 1
  • José de Jesús Olivares-Trejo
    • 1
  1. 1.Posgrado en Ciencias GenómicasUniversidad Autónoma de la Ciudad de MéxicoMéxicoMéxico

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