Wannasrichan W.Krobthong S.Morgan C.J.Armbruster E.G.Gerovac M.Yingchutrakul Y.Wongtrakoongate P.Vogel J.Aonbangkhen C.Nonejuie P.Pogliano J.Chaikeeratisak V.Mahidol University2025-03-092025-03-092025-02-01PLoS Pathogens Vol.21 No.2 (2025)15537366https://repository.li.mahidol.ac.th/handle/20.500.14594/105587Bacteriophages must hijack the gene expression machinery of their bacterial host to efficiently replicate. Recently, we have shown that the early-expressed protein gp014 of Pseudomonas nucleus-forming phage phiKZ forms a stable complex with the host ribosomes and modulates the overall protein expression profile during phage infection. Here, we discover a nucleus-forming phage, designated Churi, that is closely related to phiKZ. Churi encodes gp335, a homolog of gp014-phiKZ, which is expressed during the early stages of infection, and its overexpression in bacterial cells interferes with bacterial growth, suggesting its role in phage-host interplay. We predict experimentally that gp335 also interacts with host ribosomal proteins, similar to its homolog gp014-phiKZ, thereby strengthening its involvement in protein translation during phage infection. We further show that GFPtagged gp335 specifically localizes by clustering around the phage nucleus and remains associated with it throughout the infection cycle. The CRISPR-Cas13-mediated deletion of gp335 reveals that the mutant phage fails to replicate efficiently, resulting in an extended latent period. Altogether, our study demonstrates that gp335 is an early-expressed protein of the Chimallivirus Churi that localizes in proximity to the phage nucleus, likely serving a role in localized translation to ensure efficient phage propagation.Biochemistry, Genetics and Molecular BiologyImmunology and MicrobiologyArticleSCOPUS10.1371/journal.ppat.10129362-s2.0-8521902190815537374