Nuclear targeting activity associated with the amino terminal region of the Borna disease virus nucleoprotein.
The Borna disease virus (BDV) replicates in the nucleus. The viral p40 protein (N), which is found abundantly in the nucleus in BDV-infected cells, may play an important role in virus replication. To analyze the amino acid residues involved in the nuclear targeting of BDV N, a series of eukaryotic expression plasmids encoding deletion mutants of N was constructed and transfected into COS-7 cells. In indirect immunofluorescence assays with a rabbit anti-BDV N antiserum, wild-type N was located in the nucleus of transfected cells in the absence of other viral constituents. In contrast, mutants lacking the 13 NH2-terminal amino acid residues 1MPPKRRLVDDADA13 in common gave a cytoplasmic localization pattern. Similarly, a mutant with substitution of 4KRR6 by 4NSG6 was retained in the cytoplasm. Furthermore, a nonapeptide, 3PKRRLVDDA11, derived from the NH2-terminal region of N conferred nuclear targeting activity to beta-galactosidase, which normally resides in the cytoplasm. Thus, we have identified the nuclear targeting signal of the BDV N and narrowed it to the NH2-terminal region where 4KRR6 basic amino acid residues are located.
Kobayashi
T
T
Institute of Immunological Science, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Shoya
Y
Y
Koda
T
T
Takashima
I
I
Lai
P K
PK
Ikuta
K
K
Kakinuma
M
M
Kishi
M
M
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D001891:Borna disease virus / Q000502:physiology*; D002467:Cell Nucleus / Q000821:virology; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D017421:Sequence Analysis; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
D000956:Antigens, Viral; D014764:Viral Proteins; C077347:p40 protein, Borna disease virus