Alternate splicing in a parvoviral nonstructural gene links a common amino-terminal sequence to downstream domains which confer radically different localization and turnover characteristics.
Minute virus of mice (MVM) encodes two groups of nonstructural proteins, the 83-kDa NS-1 polypeptides encoded from a contiguous sequence in the left half of the genome and the 25-kDa NS-2 polypeptides, which share a common amino-terminal domain with NS-1 but are multiply spliced. Peptide-specific antibodies were used to demonstrate that three alternatively spliced forms of NS-2 are synthesized when synchronized A9 cells are infected with the prototype strains of MVM, MVM(p), and that each of these species migrates as two bands on sodium dodecyl sulfate-gel electrophoresis, due to the presence of both phosphorylated and unphosphorylated forms. While most NS-1 molecules are located in the nucleus, all three species of NS-2 are predominantly cytoplasmic, and their phosphorylated forms are exclusively cytoplasmic. Although both NS-1 and NS-2 molecules are synthesized early in infection, all forms of NS-2 are synthesized and accumulate three to four times as NS-1 molecules, making them the predominant virally coded proteins in the cell at this time. Despite their common amino-terminal domain, NS-2 molecules turn over rapidly while NS-1 polypeptides persist for many hours. Apart from the fact that the three NS-2 gene products are synthesized in different molar amounts, we were unable to detect any differences in the expression, stability, distribution, or phosphorylation of the various molecular forms, suggesting that these latter characteristics are mediated by their common internal exon.
Cotmore
S F
SF
Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Tattersall
P
P
D000595:Amino Acid Sequence; D000818:Animals; D016590:Aphidicolin; D001483:Base Sequence; D002213:Capsid / Q000032:analysis / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D002874:Chromosome Mapping; D004224:Diterpenes / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D007739:L Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008915:Minute Virus of Mice / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010321:Parvoviridae / Q000235:genetics*; D012326:RNA Splicing; D012367:RNA, Viral / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014758:Viral Core Proteins / Q000032:analysis / Q000235:genetics*; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins / Q000235:genetics
D004224:Diterpenes; D012367:RNA, Viral; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins; D016590:Aphidicolin