A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug
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, antisen...
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Frontiers Media S.A.
2017-11-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fcimb.2017.00492/full |
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| author | Alma Nieto David G. Pérez Ishiwara Esther Orozco Virginia Sánchez Monroy Consuelo Gómez García |
| author_facet | Alma Nieto David G. Pérez Ishiwara Esther Orozco Virginia Sánchez Monroy Consuelo Gómez García |
| author_sort | Alma Nieto |
| collection | DOAJ |
| description | 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. |
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| id | doaj.art-8d19367129484006a1e1ac5f4db8919b |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
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| last_indexed | 2024-12-23T11:59:22Z |
| publishDate | 2017-11-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-8d19367129484006a1e1ac5f4db8919b2022-12-21T17:48:01ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882017-11-01710.3389/fcimb.2017.00492301408A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine DrugAlma Nieto0David G. Pérez Ishiwara1Esther Orozco2Virginia Sánchez Monroy3Consuelo Gómez García4Laboratorio de Biomedicina Molecular I, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City, MexicoLaboratorio de Biomedicina Molecular I, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City, MexicoDepartamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico City, MexicoLaboratorio de Biomedicina Molecular I, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City, MexicoLaboratorio de Biomedicina Molecular I, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City, MexicoTranscriptional 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.http://journal.frontiersin.org/article/10.3389/fcimb.2017.00492/fullmultidrug resistanceHSEEntamoeba histolyticaEhPgp5emetinestress |
| spellingShingle | Alma Nieto David G. Pérez Ishiwara Esther Orozco Virginia Sánchez Monroy Consuelo Gómez García 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 multidrug resistance HSE Entamoeba histolytica EhPgp5 emetine stress |
| title | A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug |
| title_full | A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug |
| title_fullStr | A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug |
| title_full_unstemmed | A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug |
| title_short | A Novel Heat Shock Element (HSE) in Entamoeba histolytica that Regulates the Transcriptional Activation of the EhPgp5 Gene in the Presence of Emetine Drug |
| title_sort | novel heat shock element hse in entamoeba histolytica that regulates the transcriptional activation of the ehpgp5 gene in the presence of emetine drug |
| topic | multidrug resistance HSE Entamoeba histolytica EhPgp5 emetine stress |
| url | http://journal.frontiersin.org/article/10.3389/fcimb.2017.00492/full |
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