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. 2002 Sep;76(17):8973-6.
doi: 10.1128/jvi.76.17.8973-8976.2002.

The vaccinia virus I7L gene product is the core protein proteinase

Chelsea M Byrd  1 , Tove' C Bolken, Dennis E Hruby
Affiliations

The vaccinia virus I7L gene product is the core protein proteinase

Chelsea M Byrd et al. J Virol. 2002 Sep.

Abstract

Maturation of vaccinia virus (VV) core proteins is required for the production of infectious virions. The VV G1L and I7L gene products are the leading candidates for the viral core protein proteinase (vCPP). Using transient-expression assays, data were obtained to demonstrate that the I7L gene product and its encoded cysteine proteinase activity are responsible for vCPP activity.

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Figures

FIG. 1.
FIG. 1.
Structures of the P25K:FLAG, I7L, and G1L expression vector plasmids.
FIG. 2.
FIG. 2.
trans complementation of P25K:FLAG processing in ts16-infected cells. BSC-40 cells were infected with a temperature-sensitive VV mutant (ts16) and transfected with pRB21:17L, pRB21:G1L, p25K:FLAG, or a combination of these, and cleavage of the P25K:FLAG substrate was determined by Western blotting using anti-FLAG MAb. Lane 1, uninfected cells as a negative control; lane 2, ts16 virus alone; lane 3, ts16 with I7L; lane 4, ts16 with G1L; lane 5, ts16 with the P25K:FLAG substrate; lane 6, ts16 with I7L and P25K:FLAG; lane 7, ts16 with G1L and P25K:FLAG.
FIG. 3.
FIG. 3.
trans complementation of P25K:FLAG processing in wild-type VV-infected cells. Cells were infected with wild-type VV and transfected with pRB21:17L, p25K:FLAG, or a combination of these, and cleavage of the P25K:FLAG substrate was determined by Western blotting using anti-FLAG MAb. Lane 1, cells alone; lane 2, wild-type VV; lane 3, VV with I7L; lane 4, VV with P25K:FLAG; lane 5, VV with I7L and P25K:FLAG.
FIG. 4.
FIG. 4.
Mutational analysis of the enzyme and substrate requirements in the trans complementation of P25K:FLAG processing. Cells were infected with ts16 virus and transfected with pRB21:17L, p25K:FLAG, p25K:FLAG:IDI, p25K:FLAG:RDP, the pRB21:I7L mutant, or a combination of these, and cleavage of the P25K:FLAG substrate was determined by Western blotting using anti-FLAG MAb. Lane 1, cells alone; lane 2, ts16 alone; lane 3, ts16 with I7L; lane 4, ts16 with P25K:FLAG; lane 5 ts16 with I7L and P25K:FLAG; lane 6, ts16 with I7L and the P25K:IDI mutant; lane 7, ts16 with I7L and the P25K:RDP mutant; lane 8, ts16 with the I7L mutant and P25K:FLAG.
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