TY - JOUR

T1 - Dynamics of picornavirus RNA replication within infected cells

AU - Belsham, Graham J

AU - Normann, Preben

PY - 2008/2

Y1 - 2008/2

N2 - Replication of many picornaviruses is inhibited by low concentrations of guanidine. Guanidine-resistant mutants are readily isolated and the mutations map to the coding region for the 2C protein. Using in vitro replication assays it has been determined previously that guanidine blocks the initiation of negative-strand synthesis. We have now examined the dynamics of RNA replication, measured by quantitative RT-PCR, within cells infected with either swine vesicular disease virus (an enterovirus) or foot-and-mouth disease virus as regulated by the presence or absence of guanidine. Following the removal of guanidine from the infected cells, RNA replication occurs after a significant lag phase. This restoration of RNA synthesis requires de novo protein synthesis. Viral RNA can be maintained for at least 72 h within cells in the absence of apparent replication but guanidine-resistant virus can become predominant. Amino acid substitutions within the 2C protein that confer guanidine resistance to swine vesicular disease virus and foot-and-mouth disease virus have been identified. Even when RNA synthesis is well established, the addition of guanidine has a major impact on the level of RNA replication. Thus, the guanidine-sensitive step in RNA synthesis is important throughout the virus life cycle in cells.

AB - Replication of many picornaviruses is inhibited by low concentrations of guanidine. Guanidine-resistant mutants are readily isolated and the mutations map to the coding region for the 2C protein. Using in vitro replication assays it has been determined previously that guanidine blocks the initiation of negative-strand synthesis. We have now examined the dynamics of RNA replication, measured by quantitative RT-PCR, within cells infected with either swine vesicular disease virus (an enterovirus) or foot-and-mouth disease virus as regulated by the presence or absence of guanidine. Following the removal of guanidine from the infected cells, RNA replication occurs after a significant lag phase. This restoration of RNA synthesis requires de novo protein synthesis. Viral RNA can be maintained for at least 72 h within cells in the absence of apparent replication but guanidine-resistant virus can become predominant. Amino acid substitutions within the 2C protein that confer guanidine resistance to swine vesicular disease virus and foot-and-mouth disease virus have been identified. Even when RNA synthesis is well established, the addition of guanidine has a major impact on the level of RNA replication. Thus, the guanidine-sensitive step in RNA synthesis is important throughout the virus life cycle in cells.

KW - Carrier Proteins/physiology

KW - Cell Line

KW - Enterovirus B, Human/physiology

KW - Foot-and-Mouth Disease Virus/physiology

KW - Gene Expression Regulation, Viral

KW - Guanidine/pharmacology

KW - Picornaviridae/drug effects

KW - RNA, Viral/biosynthesis

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Viral Nonstructural Proteins/physiology

KW - Virus Replication/drug effects

U2 - 10.1099/vir.0.83385-0

DO - 10.1099/vir.0.83385-0

M3 - Journal article

C2 - 18198379

VL - 89

SP - 485

EP - 493

JO - Journal of General Virology

JF - Journal of General Virology

SN - 0022-1317

IS - Pt 2

ER -