{"doi":"10.1128/mbio.02145-23","title":"Host IP <sub>3</sub> R channels are dispensable for rotavirus Ca <sup>2+</sup> signaling but critical for intercellular Ca <sup>2+</sup> waves that prime uninfected cells for rapid virus spread","abstract":"ABSTRACT Rotavirus is a leading cause of viral gastroenteritis. A hallmark of rotavirus infection is increased cytosolic Ca 2+ caused by nonstructural protein 4 (NSP4). NSP4 is a viral ion channel that releases endoplasmic reticulum (ER) Ca 2+ , and the increased Ca 2+ signaling is critical for rotavirus replication. In addition to NSP4, host inositol 1,4,5-trisphosphate receptor (IP 3 R) ER Ca 2+ channels may contribute to rotavirus-induced Ca 2+ signaling and by extension, virus replication. Thus, we set out to determine the role of IP 3 R Ca 2+ signaling during rotavirus infection using CRISPR/Cas9 IP 3 R-knockout of MA104 cells stably expressing the GCaMP6s Ca 2+ indicator (MA104-GCaMP6s-IP 3 R-KO). Live Ca 2+ imaging showed that IP 3 R-KO did not reduce Ca 2+ signaling in infected cells but eliminated rotavirus-induced intercellular Ca 2+ waves (ICWs) and, therefore, the increased Ca 2+ signaling in surrounding, uninfected cells. MA104-GCaMP6s-IP 3 R-TKO cells showed similar rotavirus susceptibility, single-cycle replication, and viral protein expression as parental MA104-GCaMP6s cells. However, MA104-GCaMP6s-IP 3 R-TKO cells exhibited significantly smaller rotavirus plaques, decreased multi-round replication kinetics, and delayed virus spread, suggesting that rotavirus-induced ICW Ca 2+ signaling stimulates virus replication and spread. Inhibition of ICWs by blocking the purinergic receptor P2RY1 (P2Y1), which mediates the ICW Ca 2+ signals, also decreased rotavirus plaque size. Conversely, exogenous expression of P2Y1 in LLC-MK2-GCaMP6s cells, which natively lack P2Y1 and rotavirus ICWs, rescued the generation of rotavirus-induced ICWs and enabled plaque formation. In conclusion, this study shows that NSP4 Ca 2+ signals fully support rotavirus replication in individual cells; however, IP 3 R is critical for rotavirus-induced ICWs and virus spread by priming Ca 2+ -dependent pathways in surrounding cells. IMPORTANCE Many viruses exploit host Ca 2+ signaling to facilitate their replication; however, little is known about how Ca 2+ signals from different host and viral channels contribute to the overall dysregulation of Ca 2+ signaling or promote virus replication. Using cells lacking IP 3 R, a host ER Ca 2+ channel, we delineated intracellular Ca 2+ signals within virus-infected cells and intercellular Ca 2+ waves (ICWs), which increased Ca 2+ signaling in neighboring, uninfected cells. In infected cells, IP 3 R was dispensable for rotavirus-induced Ca 2+ signaling and replication, suggesting the rotavirus NSP4 viroporin supplies these signals. However, IP 3 R-mediated ICWs increase rotavirus replication kinetics and spread, indicating that the Ca 2+ signals from the ICWs may prime nearby uninfected cells to better support virus replication upon eventual infection. This “pre-emptive priming” of uninfected cells by exploiting host intercellular pathways in the vicinity of virus-infected cells represents a novel mechanism for viral reprogramming of the host to gain a replication advantage.","journal":"mBio","year":2023,"id":353462,"datarank":0.32958368660043297,"base_score":2.1972245773362196,"endowment":2.1972245773362196,"self_citation_contribution":0.32958368660043297,"citation_network_contribution":0.0,"self_endowment_contribution":0.32958368660043297,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9505,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":560910,"name":"Francesca J. Scribano","orcid":"0000-0003-2178-6324","position":1,"is_corresponding":false},{"id":1099739,"name":"J. Thomas Gebert","orcid":"0000-0001-9704-8972","position":2,"is_corresponding":false},{"id":295677,"name":"Kristen A. Engevik","orcid":"0000-0001-9807-1875","position":3,"is_corresponding":false},{"id":1100177,"name":"Jenna M. Ellis","orcid":null,"position":4,"is_corresponding":false},{"id":255993,"name":"Joseph M. Hyser","orcid":"0000-0001-5249-3482","position":5,"is_corresponding":false},{"id":295678,"name":"Jacob L. Perry","orcid":"0000-0002-0630-9266","position":0,"is_corresponding":true}],"reference_count":42,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:12:58.542950Z","pmid":"38112482","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}