Hemocyte phagosomal proteome is dynamically shaped by cytoskeleton remodeling and interorganellar communication with endoplasmic reticulum during phagocytosis in a marine invertebrate, Crassostrea gigas
Abstract Phagosomes are task-force organelles of innate immune systems, and evolutionary diversity and continuity abound in the protein machinery executing this coordinately regulated process. In order to clarify molecular mechanisms underlying phagocytosis, we studied phagocyte response to beads an...
| Published in: | Scientific Reports |
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| Main Authors: | , , , , , , , , , , , , |
| Format: | Article in Journal/Newspaper |
| Language: | English |
| Published: |
Springer Science and Business Media LLC
2020
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| Subjects: | |
| Online Access: | http://dx.doi.org/10.1038/s41598-020-63676-3 http://www.nature.com/articles/s41598-020-63676-3.pdf http://www.nature.com/articles/s41598-020-63676-3 |
| Summary: | Abstract Phagosomes are task-force organelles of innate immune systems, and evolutionary diversity and continuity abound in the protein machinery executing this coordinately regulated process. In order to clarify molecular mechanisms underlying phagocytosis, we studied phagocyte response to beads and Vibrio species, using hemocytes of the Pacific oysters ( Crassostrea gigas ) as a marine invertebrate model. Phagosomes from different stages of phagocytosis were isolated by density-gradient centrifugation, and more than 400 phagosome-associated proteins were subsequently identified via high-throughput quantitative proteomics. In modeling key networks of phagosomal proteins, our results support the essential roles of several processes driving phagosome formation and maturation, including cytoskeleton remodeling and signal transduction by Rab proteins. Several endoplasmic reticulum (ER)-associated proteins were identified, while live cell imaging confirms an apparent intimate interaction between the ER and phagosomes. In further quantitative proteomic analysis, the signal transducers Cg RhoGDI and Cg PI4K were implicated. Through experimental validation, Cg RhoGDI was shown to negatively regulate actin cytoskeleton remodeling in the formation of oyster phagosomes, while Cg PI4K signaling drives phagosome maturation and bacterial killing. Our current work illustrates the diversity and dynamic interplay of phagosomal proteins, providing a framework for better understanding host-microbe interactions during phagosome activities in under-examined invertebrate species. |
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