Correlation of proteolytic cleavage of F protein precursors in paramyxoviruses with expression of the fur, PACE4 and PC6 genes in mammalian cells.

The fusion (F) protein precursor of virulent Newcastle disease virus (NDV) strains and human parainfluenza virus type 3 (HPIV3) has a multibasic amino acid sequence at the cleavage site, and intracellular cleavage activation occurs in a variety of cells. The host protease responsible for the cleavage has been proposed to be a subtilisin-like protease (subtilase) such as furin (the product of the fur gene). We found that the lymphocyte cell lines MOLT-4, Ramos and Daudi, in addition to NALM6, lacked the ability to fully cleave the F protein precursor of virulent NDV. In contrast, MT4 as well as the non-lymphocyte cell lines HeLa and Hep2 cleaved the F protein precursor efficiently. To investigate the role of subtilases in proteolytic processing, we examined the gene expression of candidate subtilases, furin, PACE4 and PC6 in these cleavage-competent and -incompetent cells. Considerable expression of the fur gene was observed in the cleavage-competent cells, but little or no expression was detected in the cleavage-incompetent cells. PACE4 and PC6 gene expression was observed in some of the cleavage-competent cells but not in the cleavage-incompetent cells. These results suggest that furin is the protease responsible for cleavage activation of the F protein of virulent NDV strains in cultured mammalian cells and the possibility is raised that PACE4 and PC6 also participate in processing in some of the cells. On the other hand, the HPIV3 F protein was cleaved efficiently in lymphocyte cells deficient in subtilases, suggesting that an unknown protease other than furin, PACE4 or PC6 may be involved in the processing.