IDENTIFICATION OF 3 NEW JC VIRUS PROTEINS GENERATED BY ALTERNATIVE SPLICING OF THE EARLY VIRAL MESSENGER-RNA

The genome of the human polyomavirus JC Virus (JCV) encodes two regulatory proteins, large and small T antigen which are expressed early in a lytic infection, and three structural proteins, VP1, VP2, VP3, which are produced late in an infection, A fourth late protein, agnoprotein, may contribute to the assembly of the virion. In this study we demonstrate the presence of three additional early proteins, T'(135), T'(136), and T'(165), which are expressed in lytically-infected cells; T'(136) is also readily detected in JCV transformants. The three species of T' are phosphoproteins generated via an alternative splicing mechanism. This mechanism involves the excision of a second intron from the large T mRNA using a common donor splice site at JCV nucleotide 4274 and three unique acceptor splice sites at nucleotides 2918, 2777 and 2704 for T'(135), T'(136) and T'(165), respectively. The mutant JCV Delta T' was created by converting the G at nucleotide 4274 to an A, thereby disrupting the consensus sequence of the shared donor splice site without altering the amino acid sequence of any early JCV protein. Upon transfection of permissive human brain cells, JCV Delta T' replicated its DNA 10-fold less efficiently than did wild type JCV. Passage of extracts of the infected cells on to fresh human brain cells revealed that the expression of T antigen was greatly reduced and the presence of T' proteins was undetectable in the mutant versus wild type JCV-infected cells.