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Exp Parasitol. 2005 Jul;110(3):286-91. Epub 2005 Apr 9.

The Entamoeba histolytica Ehcp112 gene has a distal and weak promoter.

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1
Posgrado en Ciencias Genómicas de la Universidad Autónoma de la Ciudad de México, Mexico D.F. Ave. San Lorenzo # 290, Col. del Valle, Mexico, D.F. 03100, Mexico.

Abstract

Ehcp112 encodes the Entamoeba histolytica EhCP112 cysteine protease that is part of the EhCPADH complex. By in silico analysis we identified putative transcription factor-binding sites along 837 bp upstream the Ehcp112 gene ATG codon. A TATA-like motif (TATATAAA) was located at -36 to -29 bp, a GAAC box (GAACC) was found at -10 to -14 bp and an Inr sequence (TTCAAC) at -8 to -2 bp. These tripartite promoter elements are in non-canonical positions, downstream the transcription initiation site (-280 bp). We cloned four Ehcp112 promoter fragments in pBSCAT-ACT plasmid to obtain pI (355 bp), pII (681 bp), pIII (833 bp), and pIV (554 bp) constructs. In transfected trophozoites, only pIII drove CAT activity with 44% efficiency in relation to actin promoter activity. Our results showed the presence of a distal and weak promoter in the Ehcp112 gene. The active DNA region is inside the open reading frame of the Ehrab B gene, suggesting that expression of both genes could be coordinately regulated.

PMID:
15955326
DOI:
10.1016/j.exppara.2005.03.003
[Indexed for MEDLINE]
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