The viral protein 3CD induces an equilibrium between the viral protein and RNA synthesis in a cell-free system for poliovirus replication.
In a cell-free system derived from uninfected HeLa cells and programmed with poliovirus RNA, an unbalance between the different replication steps is observed. After programming, the vRNA is exclusively used as a template for viral translation. It takes hours before there is a switch from protein synthesis to RNA replication. This is probably the reason for the inefficient production of infectious virus (compared to the synthesis in infected cells). If, however, the cell-free system is programmed with vRNA and with a mRNA coding for the viral protein 3CD, an increase in vRNA synthesis is found early post-programming, resulting in a better balance of protein synthesis and RNA synthesis and an increased virus yield of at least 2 log10. These data show that a balance between translation, RNA replication and packaging is required to allow efficient viral proliferation. The virus yield could be increased by a further log10 by the addition of pirodavir (a capsid stabiliser) and 10% of rabbit reticulocyte lysate to the cell-free system.
Verlinden
Y
Y
Department of Microbiology and Hygiene, Vrije Universiteit Brussel, Brussels, Belgium.
Cuconati
A
A
Wimmer
E
E
Rombau
B
B
D000086042:3C Viral Proteases; D000818:Animals; D002474:Cell-Free System / Q000187:drug effects / Q000821:virology; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D010880:Piperidines / Q000494:pharmacology; D017955:Poliovirus / Q000378:metabolism*; D011724:Pyridazines / Q000494:pharmacology; D012367:RNA, Viral / Q000096:biosynthesis*; D012156:Reticulocytes; D013997:Time Factors; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
D010880:Piperidines; D011724:Pyridazines; D012367:RNA, Viral; D014764:Viral Proteins; C074589:pirodavir; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases