Mutations in the polyhedrin NLS affect the assembly and polyhedral shape of alphabaculovirus occlusion bodies

Alphabaculoviruses produce occlusion bodies (OBs) in the nucleus of the infected cells at the late stage of infection. OBs are mainly composed of a single viral protein called polyhedrin (POLH). Autographa californica multiple nucleopolyhedrovirus (AcMNPV) POLH possesses a monopartite nuclear localization signal sequence (NLS), KRKK, from 32nd to 35th residues. However, the functions of POLH NLS of other alphabaculoviruses remain unknown. Here, POLH NLS mutants of Bombyx mori nucleopolyhedrovirus (BmNPV) were generated and NLS function as well as the relationship between NLS and OB localization or morphology was investigated. Deletion or mutation of BmNPV POLH NLS severely affected POLH and OB intracellular localization. Additionally, viruses in which the arginine residue at the 33rd position of POLH was mutated produced a lower number of OBs, which was presumably due to decreased POLH accumulation in the infected cells. Furthermore, cytoplasmic OBs were morphologically aberrant, even though nuclear OB morphology was normal in the same cell. These results indicate that NLS is required for nuclear localization and efficient accumulation of BmNPV POLH, which heavily affect the number and morphology of OBs.