Expression of L* protein of Theiler's murine encephalomyelitis virus in the chronic phase of infection.

The DA strain and other members of the TO subgroup of Theiler's murine encephalomyelitis virus synthesize the L* protein from an alternative initiation codon. L* is considered to play a key role in viral persistence and demyelination in susceptible strains of mice, although this hypothesis is still controversial. By using a mutant virus that expresses FLAG epitope-tagged L*, it was demonstrated previously that L* is expressed exclusively in neurons in vivo in the acute phase of infection in the central nervous system (CNS). However, in the mutant virus, the C-H-C-C zinc-binding motif in the leader protein (L) was disrupted by the insertion of the FLAG epitope, resulting in clearance of the virus from the CNS. Therefore, a further two mutant viruses were newly generated, expressing FLAG epitope-tagged L* in which the C-H-C-C zinc-binding motif within L is spared. Both mutant viruses caused persistence and demyelination successfully in spinal cords and enabled us to identify L* immunohistochemically in the demyelinating lesions.