CRISPR screens and lectin microarrays identify high mannose N-glycan regulators
Abstract Glycans play critical roles in cellular signaling and function. Unlike proteins, glycan structures are not templated from genetic sequences but synthesized by the concerted activity of many genes, making them historically challenging to study. Here, we present a strategy that utilizes CRISP...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-11-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-53225-1 |
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| _version_ | 1849221053051568128 |
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| author | C. Kimberly Tsui Nicholas Twells Jenni Durieux Emma Doan Jacqueline Woo Noosha Khosrojerdi Janiya Brooks Ayodeji Kulepa Brant Webster Lara K. Mahal Andrew Dillin |
| author_facet | C. Kimberly Tsui Nicholas Twells Jenni Durieux Emma Doan Jacqueline Woo Noosha Khosrojerdi Janiya Brooks Ayodeji Kulepa Brant Webster Lara K. Mahal Andrew Dillin |
| author_sort | C. Kimberly Tsui |
| collection | DOAJ |
| description | Abstract Glycans play critical roles in cellular signaling and function. Unlike proteins, glycan structures are not templated from genetic sequences but synthesized by the concerted activity of many genes, making them historically challenging to study. Here, we present a strategy that utilizes CRISPR screens and lectin microarrays to uncover and characterize regulators of glycosylation. We applied this approach to study the regulation of high mannose glycans – the starting structure of all asparagine(N)-linked-glycans. We used CRISPR screens to uncover the expanded network of genes controlling high mannose levels, followed by lectin microarrays to fully measure the complex effect of select regulators on glycosylation globally. Through this, we elucidated how two high mannose regulators – TM9SF3 and the CCC complex – control complex N-glycosylation via regulating Golgi morphology and function. Notably, this allows us to interrogate Golgi function in-depth and reveals that similar disruption to Golgi morphology can lead to drastically different glycosylation outcomes. Collectively, this work demonstrates a generalizable approach for systematically dissecting the regulatory network underlying glycosylation. |
| format | Article |
| id | doaj-art-13e08ddc06d04691824a24ec2e9cac04 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-13e08ddc06d04691824a24ec2e9cac042024-11-24T12:34:24ZengNature PortfolioNature Communications2041-17232024-11-0115111310.1038/s41467-024-53225-1CRISPR screens and lectin microarrays identify high mannose N-glycan regulatorsC. Kimberly Tsui0Nicholas Twells1Jenni Durieux2Emma Doan3Jacqueline Woo4Noosha Khosrojerdi5Janiya Brooks6Ayodeji Kulepa7Brant Webster8Lara K. Mahal9Andrew Dillin10Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Chemistry, University of AlbertaDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Chemistry, University of AlbertaDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyDepartment of Chemistry, University of AlbertaDepartment of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, BerkeleyAbstract Glycans play critical roles in cellular signaling and function. Unlike proteins, glycan structures are not templated from genetic sequences but synthesized by the concerted activity of many genes, making them historically challenging to study. Here, we present a strategy that utilizes CRISPR screens and lectin microarrays to uncover and characterize regulators of glycosylation. We applied this approach to study the regulation of high mannose glycans – the starting structure of all asparagine(N)-linked-glycans. We used CRISPR screens to uncover the expanded network of genes controlling high mannose levels, followed by lectin microarrays to fully measure the complex effect of select regulators on glycosylation globally. Through this, we elucidated how two high mannose regulators – TM9SF3 and the CCC complex – control complex N-glycosylation via regulating Golgi morphology and function. Notably, this allows us to interrogate Golgi function in-depth and reveals that similar disruption to Golgi morphology can lead to drastically different glycosylation outcomes. Collectively, this work demonstrates a generalizable approach for systematically dissecting the regulatory network underlying glycosylation.https://doi.org/10.1038/s41467-024-53225-1 |
| spellingShingle | C. Kimberly Tsui Nicholas Twells Jenni Durieux Emma Doan Jacqueline Woo Noosha Khosrojerdi Janiya Brooks Ayodeji Kulepa Brant Webster Lara K. Mahal Andrew Dillin CRISPR screens and lectin microarrays identify high mannose N-glycan regulators Nature Communications |
| title | CRISPR screens and lectin microarrays identify high mannose N-glycan regulators |
| title_full | CRISPR screens and lectin microarrays identify high mannose N-glycan regulators |
| title_fullStr | CRISPR screens and lectin microarrays identify high mannose N-glycan regulators |
| title_full_unstemmed | CRISPR screens and lectin microarrays identify high mannose N-glycan regulators |
| title_short | CRISPR screens and lectin microarrays identify high mannose N-glycan regulators |
| title_sort | crispr screens and lectin microarrays identify high mannose n glycan regulators |
| url | https://doi.org/10.1038/s41467-024-53225-1 |
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