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The fushi tarazu zebra element is not required for Drosophila viability or fertility

Expression of genes in precisely controlled spatiotemporal patterns is essential for embryonic development. Much of our understanding of mechanisms regulating gene expression comes from the study of cis-regulatory elements (CREs) that direct expression of reporter genes in transgenic organisms. This...

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Autores principales: Graham, Patricia L, Fischer, Matthew D, Giri, Abhigya, Pick, Leslie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8527495/
https://www.ncbi.nlm.nih.gov/pubmed/34518886
http://dx.doi.org/10.1093/g3journal/jkab300
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author Graham, Patricia L
Fischer, Matthew D
Giri, Abhigya
Pick, Leslie
author_facet Graham, Patricia L
Fischer, Matthew D
Giri, Abhigya
Pick, Leslie
author_sort Graham, Patricia L
collection PubMed
description Expression of genes in precisely controlled spatiotemporal patterns is essential for embryonic development. Much of our understanding of mechanisms regulating gene expression comes from the study of cis-regulatory elements (CREs) that direct expression of reporter genes in transgenic organisms. This reporter-transgene approach identifies genomic regions sufficient to drive expression but fails to provide information about quantitative and qualitative contributions to endogenous expression, although such conclusions are often inferred. Here we evaluated the endogenous function of a classic Drosophila CRE, the fushi tarazu (ftz) zebra element. ftz is a pair-rule segmentation gene expressed in seven stripes during embryogenesis, necessary for formation of alternate body segments. Reporter transgenes identified the promoter-proximal zebra element as a major driver of the seven ftz stripes. We generated a precise genomic deletion of the zebra element (ftzΔZ) to assess its role in the context of native chromatin and neighboring CREs, expecting large decreases in ftz seven-stripe expression. However, significant reduction in expression was found for only one stripe, ftz stripe 4, expressed at ∼25% of wild type levels in ftzΔZ homozygotes. Defects in corresponding regions of ftzΔZ mutants suggest this level of expression borders the threshold required to promote morphological segmentation. Further, we established true-breeding lines of homozygous ftzΔZ flies, demonstrating that the body segments missing in the mutants are not required for viability or fertility. These results highlight the different types of conclusions drawn from different experimental designs and emphasize the importance of examining transcriptional regulatory mechanisms in the context of the native genomic environment.
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spelling pubmed-85274952021-10-20 The fushi tarazu zebra element is not required for Drosophila viability or fertility Graham, Patricia L Fischer, Matthew D Giri, Abhigya Pick, Leslie G3 (Bethesda) Investigation Expression of genes in precisely controlled spatiotemporal patterns is essential for embryonic development. Much of our understanding of mechanisms regulating gene expression comes from the study of cis-regulatory elements (CREs) that direct expression of reporter genes in transgenic organisms. This reporter-transgene approach identifies genomic regions sufficient to drive expression but fails to provide information about quantitative and qualitative contributions to endogenous expression, although such conclusions are often inferred. Here we evaluated the endogenous function of a classic Drosophila CRE, the fushi tarazu (ftz) zebra element. ftz is a pair-rule segmentation gene expressed in seven stripes during embryogenesis, necessary for formation of alternate body segments. Reporter transgenes identified the promoter-proximal zebra element as a major driver of the seven ftz stripes. We generated a precise genomic deletion of the zebra element (ftzΔZ) to assess its role in the context of native chromatin and neighboring CREs, expecting large decreases in ftz seven-stripe expression. However, significant reduction in expression was found for only one stripe, ftz stripe 4, expressed at ∼25% of wild type levels in ftzΔZ homozygotes. Defects in corresponding regions of ftzΔZ mutants suggest this level of expression borders the threshold required to promote morphological segmentation. Further, we established true-breeding lines of homozygous ftzΔZ flies, demonstrating that the body segments missing in the mutants are not required for viability or fertility. These results highlight the different types of conclusions drawn from different experimental designs and emphasize the importance of examining transcriptional regulatory mechanisms in the context of the native genomic environment. Oxford University Press 2021-08-26 /pmc/articles/PMC8527495/ /pubmed/34518886 http://dx.doi.org/10.1093/g3journal/jkab300 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigation
Graham, Patricia L
Fischer, Matthew D
Giri, Abhigya
Pick, Leslie
The fushi tarazu zebra element is not required for Drosophila viability or fertility
title The fushi tarazu zebra element is not required for Drosophila viability or fertility
title_full The fushi tarazu zebra element is not required for Drosophila viability or fertility
title_fullStr The fushi tarazu zebra element is not required for Drosophila viability or fertility
title_full_unstemmed The fushi tarazu zebra element is not required for Drosophila viability or fertility
title_short The fushi tarazu zebra element is not required for Drosophila viability or fertility
title_sort fushi tarazu zebra element is not required for drosophila viability or fertility
topic Investigation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8527495/
https://www.ncbi.nlm.nih.gov/pubmed/34518886
http://dx.doi.org/10.1093/g3journal/jkab300
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