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Article: Characterization of the nuclear localization signal of the borna disease virus polymerase.

Walker, Michelle Portlance / Lipkin, W Ian

Journal of virology

2001  Volume 76, Issue 16, Page(s) 8460–8467

Abstract: Borna disease virus (BDV) is a nonsegmented negative-strand RNA virus that replicates and transcribes its genome in the nucleus of infected cells. BDV proteins involved in replication and transcription must pass through the nuclear envelope to associate ... ...

Abstract Borna disease virus (BDV) is a nonsegmented negative-strand RNA virus that replicates and transcribes its genome in the nucleus of infected cells. BDV proteins involved in replication and transcription must pass through the nuclear envelope to associate with the genomic viral RNA. The RNA-dependent RNA polymerase (L) of BDV is postulated to be the catalytic enzyme of replication and transcription. We demonstrated previously that BDV L localizes to the nucleus of BDV-infected cells and L-transfected cells. Nuclear localization of the protein presupposes the presence of a nuclear localization signal (NLS) within its primary amino acid sequence or cotransport to the nucleus with another karyophilic protein. Because L localized to the nucleus in the absence of other viral proteins, we investigated the possibility that L contains an NLS. The minimal sequence required for nuclear localization of L was identified by analyzing the subcellular distribution of deletion mutants of L fused to a flag epitope tag or beta-galactosidase. Although the majority of the L fusion proteins localized to the cytoplasm of transfected BSR-T7 cells, a strong NLS (844RVVKLRIAP852) with basic and proline residues was identified. Mutation of this sequence resulted in cytoplasmic distribution of L, confirming that this sequence was necessary and sufficient to drive the nuclear localization of L.
MeSH term(s) Amino Acid Sequence ; Base Sequence ; Borna disease virus/enzymology ; Borna disease virus/genetics ; Cell Line ; Cell Nucleus/enzymology ; Cytoplasm/enzymology ; Humans ; Nuclear Localization Signals/genetics ; Peptide Mapping ; RNA Replicase/genetics ; Recombinant Fusion Proteins/genetics ; Sequence Deletion ; beta-Galactosidase/genetics
Chemical Substances Nuclear Localization Signals ; Recombinant Fusion Proteins ; RNA Replicase (EC 2.7.7.48) ; beta-Galactosidase (EC 3.2.1.23)
Language English
Publishing date 2001-05-07
Publishing country United States
Document type Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID 80174-4
ISSN 1098-5514 ; 0022-538X
ISSN (online) 1098-5514
ISSN 0022-538X
DOI 10.1128/jvi.76.16.8460-8467.2002
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