Bombyx mori nucleopolyhedrovirus FP25K is essential for maintaining a steady-state level of v-cath expression throughout the infection.
It has been previously reported that the fp25K product of Bombyx mori nucleopolyhedrovirus (BmNPV) is required for post-mortem host degradation, but the mechanism by which it regulates host degradation is still unknown. This study shows that disruption of BmNPV fp25K attenuates the expression of viral cathepsin gene (v-cath) at a late stage of infection, and thus reduces the secretion of its product V-CATH. Western blot analysis showed that secretion of V-CATH was severely reduced in BmN cells and B. mori larvae infected with Bm25KD, a BmNPV mutant lacking functional fp25K, compared to that of wild-type BmNPV. Also, reduced accumulation of pro-V-CATH in Bm25KD-infected cells was observed from 4 days postinfection (dpi), during which V-CATH was first detected in the medium of BmNPV-infected cells. qRT-PCR experiments showed that the expression levels of v-cath mRNA in wild-type- and Bm25KD-infected BmN cells were comparable at 3 dpi, but showed a marked decrease in Bm25KD-infected BmN cells at 4 dpi. Collectively, these results suggest that BmNPV FP25K is essential for the proper transcriptional regulation of v-cath and efficient secretion of V-CATH, and a steady-state level of v-cath expression during the period of V-CATH secretion (after 4 dpi) is required for post-mortem host degradation.
Katsuma
Susumu
S
Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan. katsuma@ss.ab.a.u-tokyo.ac.jp
Nakanishi
Tadashi
T
Shimada
Toru
T
D000818:Animals; D012831:Bombyx / Q000821:virology*; D002403:Cathepsins / Q000378:metabolism*; D002460:Cell Line; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D009154:Mutation; D019590:Nucleocapsid Proteins / Q000235:genetics*; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics; D014764:Viral Proteins / Q000378:metabolism*
D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D002403:Cathepsins