TY - JOUR
T1 - Entamoeba histolytica
T2 - Functional characterization of the -234 to -196 bp promoter region of the multidrug resistance EhPgp1 gene
AU - Ramirez, M. Esther
AU - Perez, D. Guillermo
AU - Nader, Elvira
AU - Gomez, Consuelo
N1 - Funding Information:
The authors thank Alfredo Padilla for his help in the artwork. This work was supported in part by the Consejo Nacional de Ciencia y Tecnología (CONACyT) and Coordinación General de Posgrado e Investigación (CGPI-IPN) (México).
PY - 2005/7
Y1 - 2005/7
N2 - The multidrug resistance EhPgp1 gene is constitutively expressed in drug resistant trophozoites from Entamoeba histolytica. It has been demonstrated that two CCAAT/enhancer binding sites located in the EhPgp1 gene promoter control its transcriptional activation. However, functional assays of the 5′ end of its promoter showed that region from -234 to -196 bp (38 bp) is also important for the EhPgp1 gene transcription. Here, we demonstrated that in the 38 bp region putative cis-activator sequences are located. In silico analysis showed the presence of GATA1, Gal4, Nit-2, and C/EBP consensus sequences. Additionally, we identified three specific DNA-protein complexes, which were competed by a C/EBP, GATA1, and HOX oligonucleotides. Finally, we partially purified three proteins of 64.4, 56.7, and 27.4 kDa. Further investigations are currently in progress to determine the identity of these nuclear factors and how they are interacting with the EhPgp1 gene promoter.
AB - The multidrug resistance EhPgp1 gene is constitutively expressed in drug resistant trophozoites from Entamoeba histolytica. It has been demonstrated that two CCAAT/enhancer binding sites located in the EhPgp1 gene promoter control its transcriptional activation. However, functional assays of the 5′ end of its promoter showed that region from -234 to -196 bp (38 bp) is also important for the EhPgp1 gene transcription. Here, we demonstrated that in the 38 bp region putative cis-activator sequences are located. In silico analysis showed the presence of GATA1, Gal4, Nit-2, and C/EBP consensus sequences. Additionally, we identified three specific DNA-protein complexes, which were competed by a C/EBP, GATA1, and HOX oligonucleotides. Finally, we partially purified three proteins of 64.4, 56.7, and 27.4 kDa. Further investigations are currently in progress to determine the identity of these nuclear factors and how they are interacting with the EhPgp1 gene promoter.
KW - Amoebiasis
KW - Drug resistance
KW - EhPgp1 gene
KW - MDR phenotype
KW - Promoter
KW - Transcription
UR - http://www.scopus.com/inward/record.url?scp=20444496845&partnerID=8YFLogxK
U2 - 10.1016/j.exppara.2005.03.011
DO - 10.1016/j.exppara.2005.03.011
M3 - Artículo
C2 - 15955318
AN - SCOPUS:20444496845
SN - 0014-4894
VL - 110
SP - 238
EP - 243
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 3 SPEC. ISS.
ER -