Characterization of a nuclear localization sequence in the polyomavirus capsid protein VP1.
Expression of VP1-beta-galactosidase fusion proteins in the yeast Saccharomyces cerevisiae was used to identify a domain of the polyomavirus VP1 capsid protein which targets this protein to the nucleus. Fusion of the first 17 amino acids of VP1 to beta-galactosidase was sufficient for nuclear localization, whereas fusion of the first 12 amino acids gave a "mixed" cytoplasmic-nuclear phenotype. Mutation of a putative targeting sequence MAPKR(5)K from R to S changed the localization of a 21 amino acid fusion protein from the nucleus to cytoplasm. These results define a nuclear location signal in the amino terminus of polyomavirus VP1 and separate this function from the high-affinity DNA binding function previously defined for this region.
Moreland
R B
RB
Division of Pediatric Oncology, Dana-Farber Cancer Institute, Children's Hospital, Boston, Massachusetts 02115.
Garcea
R L
RL
D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D036022:Capsid Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011120:Polyomavirus / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; C062481:VP1 protein, polyomavirus; D001616:beta-Galactosidase