Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis.
Patterning of C. elegans vulval cell fates relies on inductive signaling. In this induction event, a single cell, the gonadal anchor cell, secretes LIN-3/EGF and induces three out of six competent precursor cells to acquire a vulval fate. We previously showed that this developmental system is robust...
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Public Library of Science (PLoS)
2016-09-01
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| Series: | PLoS Genetics |
| Online Access: | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006278&type=printable |
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| author | Michalis Barkoulas Amhed M Vargas Velazquez Alexandre E Peluffo Marie-Anne Félix |
| author_facet | Michalis Barkoulas Amhed M Vargas Velazquez Alexandre E Peluffo Marie-Anne Félix |
| author_sort | Michalis Barkoulas |
| collection | DOAJ |
| description | Patterning of C. elegans vulval cell fates relies on inductive signaling. In this induction event, a single cell, the gonadal anchor cell, secretes LIN-3/EGF and induces three out of six competent precursor cells to acquire a vulval fate. We previously showed that this developmental system is robust to a four-fold variation in lin-3/EGF genetic dose. Here using single-molecule FISH, we find that the mean level of expression of lin-3 in the anchor cell is remarkably conserved. No change in lin-3 expression level could be detected among C. elegans wild isolates and only a low level of change-less than 30%-in the Caenorhabditis genus and in Oscheius tipulae. In C. elegans, lin-3 expression in the anchor cell is known to require three transcription factor binding sites, specifically two E-boxes and a nuclear-hormone-receptor (NHR) binding site. Mutation of any of these three elements in C. elegans results in a dramatic decrease in lin-3 expression. Yet only a single E-box is found in the Drosophilae supergroup of Caenorhabditis species, including C. angaria, while the NHR-binding site likely only evolved at the base of the Elegans group. We find that a transgene from C. angaria bearing a single E-box is sufficient for normal expression in C. elegans. Even a short 58 bp cis-regulatory fragment from C. angaria with this single E-box is able to replace the three transcription factor binding sites at the endogenous C. elegans lin-3 locus, resulting in the wild-type expression level. Thus, regulatory evolution occurring in cis within a 58 bp lin-3 fragment, results in a strict requirement for the NHR binding site and a second E-box in C. elegans. This single-cell, single-molecule, quantitative and functional evo-devo study demonstrates that conserved expression levels can hide extensive change in cis-regulatory site requirements and highlights the evolution of new cis-regulatory elements required for cell-specific gene expression. |
| format | Article |
| id | doaj-art-331b2c370b044c3ca1efd7e6d069c2f2 |
| institution | DOAJ |
| issn | 1553-7390 1553-7404 |
| language | English |
| publishDate | 2016-09-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS Genetics |
| spelling | doaj-art-331b2c370b044c3ca1efd7e6d069c2f22025-08-20T03:11:22ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042016-09-01129e100627810.1371/journal.pgen.1006278Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis.Michalis BarkoulasAmhed M Vargas VelazquezAlexandre E PeluffoMarie-Anne FélixPatterning of C. elegans vulval cell fates relies on inductive signaling. In this induction event, a single cell, the gonadal anchor cell, secretes LIN-3/EGF and induces three out of six competent precursor cells to acquire a vulval fate. We previously showed that this developmental system is robust to a four-fold variation in lin-3/EGF genetic dose. Here using single-molecule FISH, we find that the mean level of expression of lin-3 in the anchor cell is remarkably conserved. No change in lin-3 expression level could be detected among C. elegans wild isolates and only a low level of change-less than 30%-in the Caenorhabditis genus and in Oscheius tipulae. In C. elegans, lin-3 expression in the anchor cell is known to require three transcription factor binding sites, specifically two E-boxes and a nuclear-hormone-receptor (NHR) binding site. Mutation of any of these three elements in C. elegans results in a dramatic decrease in lin-3 expression. Yet only a single E-box is found in the Drosophilae supergroup of Caenorhabditis species, including C. angaria, while the NHR-binding site likely only evolved at the base of the Elegans group. We find that a transgene from C. angaria bearing a single E-box is sufficient for normal expression in C. elegans. Even a short 58 bp cis-regulatory fragment from C. angaria with this single E-box is able to replace the three transcription factor binding sites at the endogenous C. elegans lin-3 locus, resulting in the wild-type expression level. Thus, regulatory evolution occurring in cis within a 58 bp lin-3 fragment, results in a strict requirement for the NHR binding site and a second E-box in C. elegans. This single-cell, single-molecule, quantitative and functional evo-devo study demonstrates that conserved expression levels can hide extensive change in cis-regulatory site requirements and highlights the evolution of new cis-regulatory elements required for cell-specific gene expression.https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006278&type=printable |
| spellingShingle | Michalis Barkoulas Amhed M Vargas Velazquez Alexandre E Peluffo Marie-Anne Félix Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis. PLoS Genetics |
| title | Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis. |
| title_full | Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis. |
| title_fullStr | Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis. |
| title_full_unstemmed | Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis. |
| title_short | Evolution of New cis-Regulatory Motifs Required for Cell-Specific Gene Expression in Caenorhabditis. |
| title_sort | evolution of new cis regulatory motifs required for cell specific gene expression in caenorhabditis |
| url | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006278&type=printable |
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