Protein 2A is not required for Theiler's virus replication

Nonpolar mutations were introduced into all 12 regions of the genome of Theiler's murine encephalomyelitis virus. In agreement with data previously reported for other picornaviruses, mutations in regions 2B, 2C, 3A, 3B, 3C, and 3D totally abrogated viral RNA replication. Viruses with deletions in each of the capsid proteins retained RNA replication proficiency, although they were unable to propagate from cell to cell. As reported previously, mutations in the leader protein did not impair RNA replication or virus production in BHK-21 cells. Surprisingly, region 2A also appeared to be dispensable for the replication process. Indeed, up to 77 of the 133 amino acids of 2A could be deleted without significantly affecting RNA replication. 2A mutant viruses had only a slow cytopathic effect for BHK-21 cells and were totally avirulent for mice. As was the case for mutants lacking the leader protein, viruses with deletions in 2A propagated in BHK-21 cells, but their propagation was highly restricted in L929 cells.

[1]  A. C. Palmenberg,et al.  Mutational analysis of the encephalomyocarditis virus primary cleavage , 1996, Journal of virology.

[2]  J. Galama,et al.  Mengovirus leader is involved in the inhibition of host cell protein synthesis , 1996, Journal of virology.

[3]  T. Michiels,et al.  Analysis of the leader and capsid coding regions of persistent and neurovirulent strains of Theiler's virus. , 1995, Virology.

[4]  W. Kong,et al.  The leader peptide of Theiler's murine encephalomyelitis virus is a zinc-binding protein , 1995, Journal of virology.

[5]  D. Jugović,et al.  Picornavirus 2A proteinase-mediated stimulation of internal initiation of translation is dependent on enzymatic activity and the cleavage products of cellular proteins. , 1995, Virology.

[6]  H. Lipton,et al.  The leader polypeptide of Theiler's virus is essential for neurovirulence but not for virus growth in BHK cells , 1995, Journal of virology.

[7]  S. Emerson,et al.  Hepatitis A viruses with deletions in the 2A gene are infectious in cultured cells and marmosets , 1995, Journal of virology.

[8]  M. Bovee,et al.  Defective RNA replication by poliovirus mutants deficient in 2A protease cleavage activity , 1995, Journal of virology.

[9]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[10]  W. Kong,et al.  Involvement of cardiovirus leader in host cell-restricted virus expression. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[11]  D. Stone,et al.  Role for poliovirus protease 2A in cap independent translation. , 1994, The EMBO journal.

[12]  A. Paul,et al.  Studies on dicistronic polioviruses implicate viral proteinase 2Apro in RNA replication. , 1993, Virology.

[13]  P. Sarnow,et al.  Translational enhancement of the poliovirus 5' noncoding region mediated by virus-encoded polypeptide 2A. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[14]  G. Parks,et al.  Proteolytic processing of the cardioviral P2 region: primary 2A/2B cleavage in clone-derived precursors. , 1992, Virology.

[15]  M. Ryan,et al.  Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence. , 1991, The Journal of general virology.

[16]  D. Huylebroeck,et al.  In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release , 1991, Journal of virology.

[17]  M. Brahic,et al.  Genetic determinants of the demyelinating disease caused by Theiler's virus. , 1991, Microbial pathogenesis.

[18]  K. Rundell,et al.  Proteolysis at the 2A/2B junction in Theiler's murine encephalomyelitis virus. , 1991, Virology.

[19]  Macfarlane,et al.  Morphogenesis of hepatitis A virus: isolation and characterization of subviral particles , 1990, Journal of virology.

[20]  F. Tangy,et al.  Molecular cloning of the complete genome of Theiler's virus, strain DA, and production of infectious transcripts. , 1989, Microbial pathogenesis.

[21]  V. Blinov,et al.  Cysteine proteases of positive strand RNA viruses and chymotrypsin‐like serine proteases , 1989, FEBS letters.

[22]  R. Lloyd,et al.  Leader protein of foot-and-mouth disease virus is required for cleavage of the p220 component of the cap-binding protein complex , 1988, Journal of virology.

[23]  R. Lloyd,et al.  Relationship of p220 cleavage during picornavirus infection to 2A proteinase sequencing , 1988, Journal of virology.

[24]  J. Borkowski,et al.  Insights into Theiler's virus neurovirulence based on a genomic comparison of the neurovirulent GDVII and less virulent BeAn strains. , 1988, Virology.

[25]  M. Chamorro,et al.  Identification of Theiler's virus infected cells in the central nervous system of the mouse during demyelinating disease. , 1987, Microbial pathogenesis.

[26]  E. Wimmer,et al.  Poliovirus proteinase 2A induces cleavage of eucaryotic initiation factor 4F polypeptide p220 , 1987, Journal of virology.

[27]  H. Lipton,et al.  Analysis of the complete nucleotide sequence of the picornavirus Theiler's murine encephalomyelitis virus indicates that it is closely related to cardioviruses , 1987, Journal of virology.

[28]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[29]  E. Wimmer,et al.  A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein , 1986, Cell.

[30]  E. Beck,et al.  A second protease of foot-and-mouth disease virus , 1986, Journal of virology.

[31]  H. Schaller,et al.  Nucleotide sequence and genome organization of foot-and-mouth disease virus. , 1984, Nucleic acids research.

[32]  M. Janda,et al.  The nucleotide and deduced amino acid sequences of the encephalomyocarditis viral polyprotein coding region. , 1984, Nucleic acids research.

[33]  E. Ehrenfeld.  Poliovirus-induced inhibition of host-cell protein synthesis , 1982, Cell.

[34]  H. Lipton.  Theiler's virus infection in mice: an unusual biphasic disease process leading to demyelination , 1975, Infection and immunity.

[35]  A. M. Pappenheimer,et al.  OBSERVATIONS ON ENCEPHALOMYELITIS OF MICE (DA STRAIN) , 1952, The Journal of experimental medicine.

[36]  M. Theiler,et al.  ENCEPHALOMYELITIS OF MICE : I. CHARACTERISTICS AND PATHOGENESIS OF THE VIRUS. , 1940 .

[37]  M. Theiler,et al.  ENCEPHALOMYELITIS OF MICE : III. EPIDEMIOLOGY. , 1940 .

[38]  D Gani,et al.  The cleavage activities of aphthovirus and cardiovirus 2A proteins. , 1997, The Journal of general virology.

[39]  Y. Kusov,et al.  Proteinase 3C of hepatitis A virus (HAV) cleaves the HAV polyprotein P2-P3 at all sites including VP1/2A and 2A/2B. , 1994, Virology.

[40]  E. Wimmer,et al.  Genetics of poliovirus. , 1993, Annual review of genetics.

[41]  Moses Rodriguez,et al.  Theiler's murine encephalomyelitis: a model of demyelination and persistence of virus. , 1987, Critical reviews in immunology.

[42]  Thomas A. Kunkel,et al.  Rapid and efficient site-specific mutagenesis without phenotypic selection. , 1985, Proceedings of the National Academy of Sciences of the United States of America.