Hepatitis A virus polyprotein processing by Escherichia coli proteases.
Hepatitis A virus (HAV) encodes a single polyprotein, which is post-translationally processed. This processing represents an essential step in capsid formation. The virus possesses only one protease, 3C, responsible for all cleavages, except for that at the VP1/2A junction region, which is processed by cellular proteases. In this study, data demonstrates that HAV polyprotein processing by Escherichia coli protease(s) leads to the formation of particulate structures. P3 polyprotein processing in E. coli is not dependent on an active 3C protease: the same processing pattern is observed with wild-type 3C or with several 3C mutants. However, this processing pattern is temperature-dependent, since it differs at 37 or 42 degrees C. The bacterial protease(s) cleave scissile bonds other than those of HAV; this contributes to the low efficiency of particle formation.
Pintó
Rosa M
RM
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
Guix
Susana
S
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
González-Dankaart
Juan F
JF
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
Caballero
Santiago
S
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
Sánchez
Gloria
G
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
Guo
Ke-Jian
KJ
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
Ribes
Enric
E
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
Bosch
Albert
A
Department of Microbiology1 and Department of Animal and Plant Cell Biology2, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
D000086042:3C Viral Proteases; D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D030041:Hepatitis A virus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure
D011498:Protein Precursors; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases