Comprehensive interactome analysis of the spike protein of swine acute diarrhea syndrome coronavirus

•Scientific question Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a recently discovered HKU2-related bat coronavirus that causes severe and acute diarrhea and rapid weight loss in in piglets. SADS-CoV was reported to be capable of infecting cell lines derived from bats, mice, hamsters, rats, chickens, pigs, nonhuman primates, and humans. The wide range of cell tropism of SADS-CoV in vitro implies its high risk of cross-species infection.•Evidence before this study Multiple receptors for CoVs have been identified, including ACE2, DPP4 and APN. However, none of these proteins contributes to the infection of SADS-CoV.•New findings The receptor-binding domain of the SADS-CoV spike (S) protein was purified and subjected to proteomic analysis to identify the interactors of the SADS-CoV S protein. Forty-three host proteins were identified, and a Gene Ontology analysis indicated that these interactors can be grouped into categories such as “cell-cell adhesion”, “translation” “viral transcription”, suggesting that these processes may participate in the SADS-CoV life cycles. Further functional study on these interactors indicated that PPIB and vimentin can affect SADS-CoV replication.•Significance of the study This study provides an overarching view into the host interactome of the SADS-CoV S protein and highlights potential targets for the development of therapeutics against SADS-CoV. Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a recently discovered coronavirus that causes severe and acute diarrhea and rapid weight loss in piglets. SADS-CoV was reported to be capable of infecting cell lines derived from diverse species, including bats, mice, hamsters, rats, chickens, pigs, nonhuman primates, and humans, implying its high risk of cross-species infection. However, its receptor is still unknown. In this study, the receptor-binding domain of the SADS-CoV spike (S) protein was purified and then subjected to affinity purification (AP)-coupled mass spectrometry (MS)-based proteomic analysis to identify the interactors of the SADS-CoV S protein. Forty-three host proteins were identified, and a Gene Ontology analysis indicated that these interactors can be grouped into categories such as “cell-cell adhesion”, “translation” “viral transcription”, suggesting that these processes may participate in the SADS-CoV life cycles. RNA interference-based screening of these interactors indicated that PPIB and vimentin can affect SADS-CoV replication. Our study pr