Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.

Traditionally, the analysis of gene regulatory regions suffered from the caveat that it was restricted to artificial contexts (e.g. reporter constructs of limited size). With the advent of the BAC recombineering technique, genomic constructs can now be generated to test regulatory elements in their...

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Autores principales:	Lucia Gafner, Sascha Dalessi, Eliane Escher, George Pyrowolakis, Sven Bergmann, Konrad Basler
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6f6b1781f9094256890dc86a23b538de
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spelling	oai:doaj.org-article:6f6b1781f9094256890dc86a23b538de2021-11-18T09:00:35ZManipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.1932-620310.1371/journal.pone.0071224https://doaj.org/article/6f6b1781f9094256890dc86a23b538de2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23951114/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Traditionally, the analysis of gene regulatory regions suffered from the caveat that it was restricted to artificial contexts (e.g. reporter constructs of limited size). With the advent of the BAC recombineering technique, genomic constructs can now be generated to test regulatory elements in their endogenous environment. The expression of the transcriptional repressor brinker (brk) is negatively regulated by Dpp signaling. Repression is mediated by small sequence motifs, the silencer elements (SEs), that are present in multiple copies in the regulatory region of brk. In this work, we manipulated the SEs in the brk locus. We precisely quantified the effects of the individual SEs on the Brk gradient in the wing disc by employing a 1D data extraction method, followed by the quantification of the data with reference to an internal control. We found that mutating the SEs results in an expansion of the brk expression domain. However, even after mutating all predicted SEs, repression could still be observed in regions of maximal Dpp levels. Thus, our data point to the presence of additional, low affinity binding sites in the brk locus.Lucia GafnerSascha DalessiEliane EscherGeorge PyrowolakisSven BergmannKonrad BaslerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e71224 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lucia Gafner Sascha Dalessi Eliane Escher George Pyrowolakis Sven Bergmann Konrad Basler Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
description	Traditionally, the analysis of gene regulatory regions suffered from the caveat that it was restricted to artificial contexts (e.g. reporter constructs of limited size). With the advent of the BAC recombineering technique, genomic constructs can now be generated to test regulatory elements in their endogenous environment. The expression of the transcriptional repressor brinker (brk) is negatively regulated by Dpp signaling. Repression is mediated by small sequence motifs, the silencer elements (SEs), that are present in multiple copies in the regulatory region of brk. In this work, we manipulated the SEs in the brk locus. We precisely quantified the effects of the individual SEs on the Brk gradient in the wing disc by employing a 1D data extraction method, followed by the quantification of the data with reference to an internal control. We found that mutating the SEs results in an expansion of the brk expression domain. However, even after mutating all predicted SEs, repression could still be observed in regions of maximal Dpp levels. Thus, our data point to the presence of additional, low affinity binding sites in the brk locus.
format	article
author	Lucia Gafner Sascha Dalessi Eliane Escher George Pyrowolakis Sven Bergmann Konrad Basler
author_facet	Lucia Gafner Sascha Dalessi Eliane Escher George Pyrowolakis Sven Bergmann Konrad Basler
author_sort	Lucia Gafner
title	Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
title_short	Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
title_full	Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
title_fullStr	Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
title_full_unstemmed	Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
title_sort	manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/6f6b1781f9094256890dc86a23b538de
work_keys_str_mv	AT luciagafner manipulatingthesensitivityofsignalinducedrepressionquantificationandconsequencesofalteredbrinkergradients AT saschadalessi manipulatingthesensitivityofsignalinducedrepressionquantificationandconsequencesofalteredbrinkergradients AT elianeescher manipulatingthesensitivityofsignalinducedrepressionquantificationandconsequencesofalteredbrinkergradients AT georgepyrowolakis manipulatingthesensitivityofsignalinducedrepressionquantificationandconsequencesofalteredbrinkergradients AT svenbergmann manipulatingthesensitivityofsignalinducedrepressionquantificationandconsequencesofalteredbrinkergradients AT konradbasler manipulatingthesensitivityofsignalinducedrepressionquantificationandconsequencesofalteredbrinkergradients
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Manipulating the sensitivity of signal-induced repression: quantification and consequences of altered brinker gradients.

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