A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug

Frontiers in Cellular and Infection Microbiology

Gó¹  Orozco, Esther²  Nieto, Alma²  Pé³  Sá³  nchez Monroy, Virginia⁴  rez Ishiwara, David G.⁴  mez Garcí⁵  a, Consuelo⁷ 

[1]Departamento de Infectó

[2]Laboratorio de Biomedicina Molecular I, Escuela Nacional de Medicina y Homeopatí

[3]a, Instituto Polité

[4]cnico Nacional, Mexico

[5]mica y Patogé

[6]n y de Estudios Avanzados del Instituto Polité

[7]nesis Molecular, Centro de Investigació

关键词: multidrug resistance;    hse;    Entamoeba histolytica;    EhPgp5;    Emetine;    stress;   

DOI  :  10.3389/fcimb.2017.00492

学科分类：生物科学（综合）

来源: Frontiers

【 摘 要 】

ABSTRACT 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.

【 授权许可】

CC BY