TY - JOUR
T1 - A novel heat shock element (HSE) in Entamoeba histolytica that regulates the transcriptional activation of the EhPgp5 gene in the presence of emetine drug
AU - Nieto, Alma
AU - Ishiwara, David G.Pérez
AU - Orozco, Esther
AU - Sánchez Monroy, Virginia
AU - Gómez García, Consuelo
N1 - Publisher Copyright:
© 2017 Nieto, Pérez Ishiwara, Orozco, Sánchez Monroy and Gómez García.
PY - 2017/11/29
Y1 - 2017/11/29
N2 - Transcriptional regulation of the multidrug resistance EhPgp5 gene in Entamoeba histolytica is induced by emetine stress. EhPgp5 overexpression alters the chloride-dependent currents that cause trophozoite swelling, diminishing induced programmed cell death (PCD) susceptibility. In contrast, antisense inhibition of P-glycoprotein (PGP) expression produces synchronous death of trophozoites and the enhancement of the biochemical and morphological characteristics of PCD induced by G418. Transcriptional gene regulation analysis identified a 59 bp region at position -170 to -111 bp promoter as putative emetine response elements (EREs). However, insights into transcription factors controlling EhPgp5 gene transcription are missing; to fill this knowledge gap, we used deletion studies and transient CAT activity assays. Our findings suggested an activating motif (-151 to -136 bp) that corresponds to a heat shock element (HSE). Gel-shift assays, UV-crosslinking, binding protein purification, and western blotting assays revealed proteins of 94, 66, 62, and 51 kDa binding to the EhPgp5 HSE that could be heat shock-like transcription factors that regulate the transcriptional activation of the EhPgp5 gene in the presence of emetine drug.
AB - Transcriptional regulation of the multidrug resistance EhPgp5 gene in Entamoeba histolytica is induced by emetine stress. EhPgp5 overexpression alters the chloride-dependent currents that cause trophozoite swelling, diminishing induced programmed cell death (PCD) susceptibility. In contrast, antisense inhibition of P-glycoprotein (PGP) expression produces synchronous death of trophozoites and the enhancement of the biochemical and morphological characteristics of PCD induced by G418. Transcriptional gene regulation analysis identified a 59 bp region at position -170 to -111 bp promoter as putative emetine response elements (EREs). However, insights into transcription factors controlling EhPgp5 gene transcription are missing; to fill this knowledge gap, we used deletion studies and transient CAT activity assays. Our findings suggested an activating motif (-151 to -136 bp) that corresponds to a heat shock element (HSE). Gel-shift assays, UV-crosslinking, binding protein purification, and western blotting assays revealed proteins of 94, 66, 62, and 51 kDa binding to the EhPgp5 HSE that could be heat shock-like transcription factors that regulate the transcriptional activation of the EhPgp5 gene in the presence of emetine drug.
KW - EhPgp5
KW - Emetine
KW - Entamoeba histolytica
KW - HSE
KW - Multidrug resistance
KW - Stress
UR - http://www.scopus.com/inward/record.url?scp=85043778308&partnerID=8YFLogxK
U2 - 10.3389/fcimb.2017.00492
DO - 10.3389/fcimb.2017.00492
M3 - Artículo
C2 - 29238701
SN - 2235-2988
VL - 7
JO - Frontiers in Cellular and Infection Microbiology
JF - Frontiers in Cellular and Infection Microbiology
IS - NOV
M1 - 492
ER -