Kinetochore Proteins CENPB and LIC2 are critical for Episome Persistence and Replication of Kaposis's Sarcoma-Associated Herpesvirus

Kaposi's sarcoma-associated herpesvirus (KSHV) establishes lifelong latency by maintaining its genome as circular episomes tethered to host chromatin and faithfully segregated during mitosis. This process is primarily mediated by the latency-associated nuclear antigen (LANA), which interacts with host chromosome-associated factors to ensure viral genome persistence. Several kinetochore proteins, including CENP-F, Bub1, BubR1, Aurora B, Survivin, and NuMA, have been implicated in this process, yet the broader contribution of kinetochore components remains poorly defined. We performed a systematic shRNA-based screen of the 37 host genes encoding kinetochore-associated proteins to evaluate their role in KSHV episome maintenance. We validated involvement of previously reported factors and identified Centromere Protein B (CENP-B) and Light Intermediate Chain 2 of the dynein complex (LIC2) as novel kinetochore-associated proteins critical for KSHV episome persistence. Loss of these proteins significantly reduced episome stability, highlighting previously unrecognized host-virus interactions at the chromosome segregation machinery. These findings identify critical host factors that expand the repertoire of kinetochore proteins co-opted by KSHV for episome maintenance and provide new mechanistic insights into how viral episomes exploit host chromosome segregation pathways for long-term persistence. Our work underscores kinetochore proteins as a broader class of host dependency factors in gamma-herpesvirus biology and suggests potential therapeutic targets for KSHV-associated malignancies.