biostudies-literatureUnknown28(11)Gupta DLisH-domain-containing proteins are involved in diverse cellular processes and disease mechanisms, yet their functional interaction landscape remains poorly characterized. Here, we employed a proteomics-based strategy to systematically map the interaction network of 27 LisH-domain-containing proteins, uncovering 1,410 high-confidence interactions-90% of which are previously unreported. This network reveals unanticipated roles for LisH proteins in cellular regulation and uncovers links to human disease, including cancer-associated interactions. Focusing on Lis1, a cytoplasmic dynein regulator, we identify the RBR-E3 ubiquitin ligase ARIH2 as a key functional interactor. We show that Lis1 promotes ARIH2 deneddylation via the COP9 signalosome (CSN), modulating its ubiquitin ligase activity. AiScience113912https://www.ebi.ac.uk/biostudies/studies/S-EPMC12648486biostudies-literatureA LisH-domain protein interaction map reveals a Lis1-ARIH2-dynein regulatory axis.PMC12648486Gupta DMaddika SfalseA LisH-domain protein interaction map reveals a Lis1-ARIH2-dynein regulatory axis.LisH-domain-containing proteins are involved in diverse cellular processes and disease mechanisms, yet their functional interaction landscape remains poorly characterized. Here, we employed a proteomics-based strategy to systematically map the interaction network of 27 LisH-domain-containing proteins, uncovering 1,410 high-confidence interactions-90% of which are previously unreported. This network reveals unanticipated roles for LisH proteins in cellular regulation and uncovers links to human disease, including cancer-associated interactions. Focusing on Lis1, a cytoplasmic dynein regulator, we identify the RBR-E3 ubiquitin ligase ARIH2 as a key functional interactor. We show that Lis1 promotes ARIH2 deneddylation via the COP9 signalosome (CSN), modulating its ubiquitin ligase activity. A2025-01-01T00:00:00Z2025 Nov2026-06-05T18:49:43.792Z2026-05-20T03:13:38.442ZS-EPMC126484864131238610.1016/j.isci.2025.113912