ESCRT ‐dependent protein sorting is required for the viability of yeast clathrin‐mediated endocytosis mutants
| العنوان: | |
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| المؤلفون: | Derek C. Prosser, Yorke Zhang, Beverly Wendland, Samantha Y. Lux, Pedro G. Castiñeira, Joanna E. Poprawski, Tony Yao, Carolyn R. Norris, Kyle Hoban, Sanjana Pesari, James Shepherdson |
| المصدر: | Traffic. 21:430-450 |
| بيانات النشر: | Wiley, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Saccharomyces cerevisiae Proteins, Epsin, Endosome, Endocytic cycle, Endocytic recycling, Endocytosis Pathway, Saccharomyces cerevisiae, macromolecular substances, Endocytosis, Biochemistry, Clathrin, 03 medical and health sciences, 0302 clinical medicine, Structural Biology, Gene Expression Regulation, Fungal, Genetics, Molecular Biology, 030304 developmental biology, Adenosine Triphosphatases, 0303 health sciences, Endosomal Sorting Complexes Required for Transport, biology, Cell Biology, Cell biology, Protein Transport, biology.protein, Ap180, 030217 neurology & neurosurgery |
| الوصف: | Endocytosis regulates many processes, including signaling pathways, nutrient uptake, and protein turnover. During clathrin-mediated endocytosis (CME), adaptors bind to cytoplasmic regions of transmembrane cargo proteins, and many endocytic adaptors are also directly involved in the recruitment of clathrin. This clathrin-associated sorting protein family includes the yeast epsins, Ent1/2, and AP180/PICALM homologs, Yap1801/2. Mutant strains lacking these four adaptors, but expressing an epsin N-terminal homology (ENTH) domain necessary for viability (4Δ+ENTH), exhibit endocytic defects, such as cargo accumulation at the plasma membrane (PM). This CME-deficient strain provides a sensitized background ideal for revealing cellular components that interact with clathrin adaptors. We performed a mutagenic screen to identify alleles that are lethal in 4Δ+ENTH cells using a colony-sectoring reporter assay. After isolating candidate synthetic lethal genes by complementation, we confirmed that mutations in VPS4 led to inviability of a 4Δ+ENTH strain. Vps4 mediates the final step of endosomal sorting complex required for transport (ESCRT)-dependent trafficking, and we found that multiple ESCRTs are also essential in 4Δ+ENTH cells, including Snf7, Snf8 and Vps36. Deletion of VPS4 from an end3Δ strain, another CME mutant, similarly resulted in inviability, and upregulation of a clathrin-independent endocytosis pathway rescued 4Δ+ENTH vps4Δ cells. Loss of Vps4 from an otherwise wild-type background caused multiple cargoes to accumulate at the PM because of an increase in Rcy1-dependent recycling of internalized protein to the cell surface. Additionally, vps4Δ rcy1Δ mutants exhibited deleterious growth phenotypes. Together, our findings reveal previously unappreciated effects of disrupted ESCRT-dependent trafficking on endocytic recycling and the PM. |
| تدمد: | 1600-0854 1398-9219 |
| DOI: | 10.1111/tra.12731 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7b1f4bb498247976315f85258e9311bd https://doi.org/10.1111/tra.12731 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....7b1f4bb498247976315f85258e9311bd |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16000854 13989219 |
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| DOI: | 10.1111/tra.12731 |