Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins
Summary: Tetraspanins, a superfamily of membrane proteins, mediate diverse biological processes through tetraspanin-enriched microdomains in the plasma membrane. However, how their cell-surface presentation is controlled remains unclear. To identify the regulators of tetraspanin trafficking, we cond...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-02-01
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| Series: | Cell Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124723000761 |
| _version_ | 1797936381788422144 |
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| author | Jicheng Yang Fusheng Guo Hui San Chin Gao Bin Chen Chow Hiang Ang Qingsong Lin Wanjin Hong Nai Yang Fu |
| author_facet | Jicheng Yang Fusheng Guo Hui San Chin Gao Bin Chen Chow Hiang Ang Qingsong Lin Wanjin Hong Nai Yang Fu |
| author_sort | Jicheng Yang |
| collection | DOAJ |
| description | Summary: Tetraspanins, a superfamily of membrane proteins, mediate diverse biological processes through tetraspanin-enriched microdomains in the plasma membrane. However, how their cell-surface presentation is controlled remains unclear. To identify the regulators of tetraspanin trafficking, we conduct sequential genome-wide loss-of-function CRISPR-Cas9 screens based on cell-surface expression of a tetraspanin member, TSPAN8. Several genes potentially involved in endoplasmic reticulum (ER) targeting, different biological processes in the Golgi apparatus, and protein trafficking are identified and functionally validated. Importantly, we find that biantennary N-glycans generated by MGAT1/2, but not more complex glycan structures, are important for cell-surface tetraspanin expression. Moreover, we unravel that SPPL3, a Golgi intramembrane-cleaving protease reported previously to act as a sheddase of multiple glycan-modifying enzymes, controls cell-surface tetraspanin expression through a mechanism associated with lacto-series glycolipid biosynthesis. Our study provides critical insights into the molecular regulation of cell-surface presentation of tetraspanins with implications for strategies to manipulate their functions, including cancer cell invasion. |
| first_indexed | 2024-04-10T18:29:58Z |
| format | Article |
| id | doaj.art-85cf40776e8e4f449d0d383fbf30fcde |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-04-10T18:29:58Z |
| publishDate | 2023-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-85cf40776e8e4f449d0d383fbf30fcde2023-02-02T04:48:19ZengElsevierCell Reports2211-12472023-02-01422112065Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspaninsJicheng Yang0Fusheng Guo1Hui San Chin2Gao Bin Chen3Chow Hiang Ang4Qingsong Lin5Wanjin Hong6Nai Yang Fu7Cancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore 169857, SingaporeCancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore 169857, SingaporeCancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore 169857, SingaporeCancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore 169857, SingaporeCancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore 169857, SingaporeDepartment of Biological Sciences, National University of Singapore, Singapore 117543, SingaporeInstitute of Molecular and Cell Biology, Agency for Science, Technology, and Research (A∗STAR), Singapore 138673, SingaporeCancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore 169857, Singapore; Department of Physiology, National University of Singapore, Singapore 117593, Singapore; Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medicine, University of Melbourne, Parkville, VIC 3010, Australia; Corresponding authorSummary: Tetraspanins, a superfamily of membrane proteins, mediate diverse biological processes through tetraspanin-enriched microdomains in the plasma membrane. However, how their cell-surface presentation is controlled remains unclear. To identify the regulators of tetraspanin trafficking, we conduct sequential genome-wide loss-of-function CRISPR-Cas9 screens based on cell-surface expression of a tetraspanin member, TSPAN8. Several genes potentially involved in endoplasmic reticulum (ER) targeting, different biological processes in the Golgi apparatus, and protein trafficking are identified and functionally validated. Importantly, we find that biantennary N-glycans generated by MGAT1/2, but not more complex glycan structures, are important for cell-surface tetraspanin expression. Moreover, we unravel that SPPL3, a Golgi intramembrane-cleaving protease reported previously to act as a sheddase of multiple glycan-modifying enzymes, controls cell-surface tetraspanin expression through a mechanism associated with lacto-series glycolipid biosynthesis. Our study provides critical insights into the molecular regulation of cell-surface presentation of tetraspanins with implications for strategies to manipulate their functions, including cancer cell invasion.http://www.sciencedirect.com/science/article/pii/S2211124723000761CP: Cell biology |
| spellingShingle | Jicheng Yang Fusheng Guo Hui San Chin Gao Bin Chen Chow Hiang Ang Qingsong Lin Wanjin Hong Nai Yang Fu Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins Cell Reports CP: Cell biology |
| title | Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins |
| title_full | Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins |
| title_fullStr | Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins |
| title_full_unstemmed | Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins |
| title_short | Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins |
| title_sort | sequential genome wide crispr cas9 screens identify genes regulating cell surface expression of tetraspanins |
| topic | CP: Cell biology |
| url | http://www.sciencedirect.com/science/article/pii/S2211124723000761 |
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