Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila
A BMP gradient is essential for patterning the dorsal-ventral axis of invertebrate and vertebrate embryos. The extracellular BMP binding protein Short Gastrulation (Sog) in Drosophila plays a key role in BMP gradient formation. In this study, we combine genome editing, structural and developmental a...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2022-06-01
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| Series: | Biology Open |
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| Online Access: | http://bio.biologists.org/content/11/6/bio059199 |
| _version_ | 1818488574265262080 |
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| author | Sophie L. Frampton Catherine Sutcliffe Clair Baldock Hilary L. Ashe |
| author_facet | Sophie L. Frampton Catherine Sutcliffe Clair Baldock Hilary L. Ashe |
| author_sort | Sophie L. Frampton |
| collection | DOAJ |
| description | A BMP gradient is essential for patterning the dorsal-ventral axis of invertebrate and vertebrate embryos. The extracellular BMP binding protein Short Gastrulation (Sog) in Drosophila plays a key role in BMP gradient formation. In this study, we combine genome editing, structural and developmental approaches to study Sog function in Drosophila. We generate a sog knockout fly stock, which allows simple reintegration of altered versions of the sog coding sequence. As proof-of-principle, we test the requirement for two cysteine residues that were previously identified as targets for palmitoylation, which has been proposed to enhance Sog secretion. However, we show that the sogC27,28S mutant is viable with only very mild phenotypes, indicating that these residues and their potential modification are not critical for Sog secretion in vivo. Additionally, we use experimental negative stain EM imaging and hydrodynamic data to validate the AlphaFold structure prediction for Sog. The model suggests a more compact shape than the vertebrate ortholog Chordin and conformational flexibility between the C-terminal von Willebrand C domains. We discuss how this altered compactness may contribute to mechanistic differences in Sog and Chordin function during BMP gradient formation. This article has an associated First Person interview with the first author of the paper. |
| first_indexed | 2024-12-10T16:52:58Z |
| format | Article |
| id | doaj.art-e7613e35f19846d9b444ff1a20ef6d68 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-10T16:52:58Z |
| publishDate | 2022-06-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-e7613e35f19846d9b444ff1a20ef6d682022-12-22T01:40:51ZengThe Company of BiologistsBiology Open2046-63902022-06-0111610.1242/bio.059199059199Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in DrosophilaSophie L. Frampton0Catherine Sutcliffe1Clair Baldock2Hilary L. Ashe3 Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK Wellcome Centre for Cell-Matrix Research, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, M13 9PT, UK Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK A BMP gradient is essential for patterning the dorsal-ventral axis of invertebrate and vertebrate embryos. The extracellular BMP binding protein Short Gastrulation (Sog) in Drosophila plays a key role in BMP gradient formation. In this study, we combine genome editing, structural and developmental approaches to study Sog function in Drosophila. We generate a sog knockout fly stock, which allows simple reintegration of altered versions of the sog coding sequence. As proof-of-principle, we test the requirement for two cysteine residues that were previously identified as targets for palmitoylation, which has been proposed to enhance Sog secretion. However, we show that the sogC27,28S mutant is viable with only very mild phenotypes, indicating that these residues and their potential modification are not critical for Sog secretion in vivo. Additionally, we use experimental negative stain EM imaging and hydrodynamic data to validate the AlphaFold structure prediction for Sog. The model suggests a more compact shape than the vertebrate ortholog Chordin and conformational flexibility between the C-terminal von Willebrand C domains. We discuss how this altered compactness may contribute to mechanistic differences in Sog and Chordin function during BMP gradient formation. This article has an associated First Person interview with the first author of the paper.http://bio.biologists.org/content/11/6/bio059199drosophilashort gastrulationcrispr-cas9alphafoldmodelpalmitoylation |
| spellingShingle | Sophie L. Frampton Catherine Sutcliffe Clair Baldock Hilary L. Ashe Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila Biology Open drosophila short gastrulation crispr-cas9 alphafold model palmitoylation |
| title | Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila |
| title_full | Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila |
| title_fullStr | Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila |
| title_full_unstemmed | Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila |
| title_short | Modelling the structure of Short Gastrulation and generation of a toolkit for studying its function in Drosophila |
| title_sort | modelling the structure of short gastrulation and generation of a toolkit for studying its function in drosophila |
| topic | drosophila short gastrulation crispr-cas9 alphafold model palmitoylation |
| url | http://bio.biologists.org/content/11/6/bio059199 |
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