@article {68316,
	title = {The SARS-CoV-2 accessory protein Orf3a is not an ion channel, but does interact with trafficking proteins},
	journal = {eLife},
	year = {2023},
	month = {2023 Jan 25},
	abstract = {<p>The severe acute respiratory syndrome associated coronavirus 2 (SARS-CoV-2) and SARS-CoV-1 accessory protein Orf3a colocalizes with markers of the plasma membrane, endocytic pathway, and Golgi apparatus. Some reports have led to annotation of both Orf3a proteins as a viroporin. Here we show that neither SARS-CoV-2 nor SARS-CoV-1 form functional ion conducting pores and that the conductances measured are common contaminants in overexpression and with high levels of protein in reconstitution studies. Cryo-EM structures of both SARS-CoV-2 and SARS-CoV-1 Orf3a display a narrow constriction and the presence of a basic aqueous vestibule, which would not favor cation permeation. We observe enrichment of the late endosomal marker Rab7 upon SARS-CoV-2 Orf3a overexpression, and co-immunoprecipitation with VPS39. Interestingly, SARS-CoV-1 Orf3a does not cause the same cellular phenotype as SARS-CoV-2 Orf3a and does not interact with VPS39. To explain this difference, we find that a divergent, unstructured loop of SARS-CoV-2 Orf3a facilitates its binding with VPS39, a HOPS complex tethering protein involved in late endosome and autophagosome fusion with lysosomes. We suggest that the added loop enhances SARS-CoV-2 Orf3a ability to co-opt host cellular trafficking mechanisms for viral exit or host immune evasion.</p>
},
	doi = {10.7554/eLife.84477},
	url = {https://doi.org/10.7554/eLife.84477},
	author = {Alexandria N. Miller and Patrick R. Houlihan and Ella Matamala and Deny Cabezas-Bratesco and Gi Young Lee and Ben Cristofori-Armstrong and Tanya L. Dilan and Silvia Sanchez-Martinez and Doreen Matthies and Rui Yan and Zhiheng Yu and Dejian Ren and Sebastian E. Brauchi and David E. Clapham}
}
