Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila

Abstract Homeoproteins contain the conserved homeodomain (HD) and have an important role determining embryo body plan during development. HDs increase their DNA-binding specificity by interacting with additional cofactors outlining a Hox interactome with a multiplicity of protein-protein interactio...

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Autores principales: Singh, Amit, Altamirano Torres, Claudia Dalila, Salinas Hernández, Jannet Edith, Cárdenas Chávez, Diana Linda, Rodríguez Padilla, Cristina, Reséndez Pérez, Diana
Formato: Artículo
Lenguaje:inglés
Publicado: 2018
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Acceso en línea:http://eprints.uanl.mx/16086/1/64.pdf
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author Singh, Amit
Altamirano Torres, Claudia Dalila
Salinas Hernández, Jannet Edith
Cárdenas Chávez, Diana Linda
Rodríguez Padilla, Cristina
Reséndez Pérez, Diana
author_facet Singh, Amit
Altamirano Torres, Claudia Dalila
Salinas Hernández, Jannet Edith
Cárdenas Chávez, Diana Linda
Rodríguez Padilla, Cristina
Reséndez Pérez, Diana
author_sort Singh, Amit
collection Repositorio Institucional
description Abstract Homeoproteins contain the conserved homeodomain (HD) and have an important role determining embryo body plan during development. HDs increase their DNA-binding specificity by interacting with additional cofactors outlining a Hox interactome with a multiplicity of protein-protein interactions. In Drosophila, the first link of functional contact with a general transcription factor (GTF) was found between Antennapedia (Antp) and BIP2 (TFIID complex). Hox proteins also interact with other components of Pol II machinery such as the subunit Med19 from Mediator (MED) complex, TFIIEβand transcription-pausing factor M1BP. All these interactions clearly demonstrate Hox-driven transcriptional regulation, but the precise molecular mechanism remains unclear. In this paper, we focused on the Antp-TFIIEβ protein-protein interface to establish the specific contacts as well as its functional role. Using Bimolecular Fluorescence Complementation (BiFC) in cell culture and in vivo we found that TFIIEβinteracts with Antp through the HD independently of the YPWM motif and the direct physical interaction is at helix 2, specifically aminoacidic positions I32 and H36 of Antp. We also found, through ectopic assays, that these two positions in helix 2 are crucial for Antp homeotic function in head involution, and thoracic and antenna-to tarsus transformations. Interestingly, overexpression of Antp and TFIIEβin the antennal disc showed that this interaction is required for the antenna-to-tarsus transformation. In conclusion, interaction of Antp with TFIIEβis important for the functional specificity of Antennapedia, and amino acids 32 and 36 in Antp HD helix 2 are key for this interaction. Our results open the possibility to more broadly analyze Antp-TFIIEβinteraction on the transcriptional control for the activation and/ or repression of target genes in the Hox interactome during Drosophila development.
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spelling eprints-160862025-07-18T17:18:47Z http://eprints.uanl.mx/16086/ Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila Singh, Amit Altamirano Torres, Claudia Dalila Salinas Hernández, Jannet Edith Cárdenas Chávez, Diana Linda Rodríguez Padilla, Cristina Reséndez Pérez, Diana QR Microbiología Abstract Homeoproteins contain the conserved homeodomain (HD) and have an important role determining embryo body plan during development. HDs increase their DNA-binding specificity by interacting with additional cofactors outlining a Hox interactome with a multiplicity of protein-protein interactions. In Drosophila, the first link of functional contact with a general transcription factor (GTF) was found between Antennapedia (Antp) and BIP2 (TFIID complex). Hox proteins also interact with other components of Pol II machinery such as the subunit Med19 from Mediator (MED) complex, TFIIEβand transcription-pausing factor M1BP. All these interactions clearly demonstrate Hox-driven transcriptional regulation, but the precise molecular mechanism remains unclear. In this paper, we focused on the Antp-TFIIEβ protein-protein interface to establish the specific contacts as well as its functional role. Using Bimolecular Fluorescence Complementation (BiFC) in cell culture and in vivo we found that TFIIEβinteracts with Antp through the HD independently of the YPWM motif and the direct physical interaction is at helix 2, specifically aminoacidic positions I32 and H36 of Antp. We also found, through ectopic assays, that these two positions in helix 2 are crucial for Antp homeotic function in head involution, and thoracic and antenna-to tarsus transformations. Interestingly, overexpression of Antp and TFIIEβin the antennal disc showed that this interaction is required for the antenna-to-tarsus transformation. In conclusion, interaction of Antp with TFIIEβis important for the functional specificity of Antennapedia, and amino acids 32 and 36 in Antp HD helix 2 are key for this interaction. Our results open the possibility to more broadly analyze Antp-TFIIEβinteraction on the transcriptional control for the activation and/ or repression of target genes in the Hox interactome during Drosophila development. 2018-10-15 Article PeerReviewed text en cc_by_nc_nd http://eprints.uanl.mx/16086/1/64.pdf http://eprints.uanl.mx/16086/1.haspreviewThumbnailVersion/64.pdf Singh, Amit y Altamirano Torres, Claudia Dalila y Salinas Hernández, Jannet Edith y Cárdenas Chávez, Diana Linda y Rodríguez Padilla, Cristina y Reséndez Pérez, Diana (2018) Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila. PLOS ONE, 13 (10). pp. 1-14. ISSN 1932-6203 http://doi.org/10.1371/journal.pone.0205905 doi:10.1371/journal.pone.0205905
spellingShingle QR Microbiología
Singh, Amit
Altamirano Torres, Claudia Dalila
Salinas Hernández, Jannet Edith
Cárdenas Chávez, Diana Linda
Rodríguez Padilla, Cristina
Reséndez Pérez, Diana
Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila
thumbnail https://rediab.uanl.mx/themes/sandal5/images/online.png
title Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila
title_full Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila
title_fullStr Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila
title_full_unstemmed Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila
title_short Transcription factor TFIIEβ interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila
title_sort transcription factor tfiieβ interacts with two exposed positions in helix 2 of the antennapedia homeodomain to control homeotic function in drosophila
topic QR Microbiología
url http://eprints.uanl.mx/16086/1/64.pdf
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