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Journal of Virology, October 2001, p. 9393-9406, Vol. 75, No. 19
Department of Plant and Microbial Biology,
University of California, Berkeley, California 94720
Received 6 April 2001/Accepted 21 June 2001
We have characterized the interaction and nuclear localization of
the nucleocapsid (N) protein and phosphoprotein (P) of sonchus yellow
net nucleorhabdovirus. Expression studies with plant and yeast cells
revealed that both N and P are capable of independent nuclear import.
Site-specific mutagenesis and deletion analyses demonstrated that N
contains a carboxy-terminal bipartite nuclear localization signal (NLS)
located between amino acids 465 and 481 and that P contains a
karyophillic region between amino acids 40 and 124. The N NLS was fully
capable of functioning outside of the context of the N protein and was
able to direct the nuclear import of a synthetic protein fusion
consisting of green fluorescent protein fused to glutathione
S-transferase (GST). Expression and mapping studies
suggested that the karyophillic domain in P is located within the
N-binding domain. Coexpression of N and P drastically affected their
localization patterns relative to those of individually expressed
proteins and resulted in a shift of both proteins to a subnuclear
region. Yeast two-hybrid and GST pulldown experiments verified the N-P
and P-P interactions, and deletion analyses have identified the N and P
interacting domains. N NLS mutants were not transported to the nucleus
by import-competent P, presumably because N binding masks the P NLS.
Taken together, our results support a model for independent entry of N
and P into the nucleus followed by associations that mediate subnuclear localization.
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.19.9393-9406.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Interactions and Nuclear Import of the N and P Proteins of
Sonchus Yellow Net Virus, a Plant Nucleorhabdovirus
*
Corresponding author. Mailing address: Department of
Plant and Microbial Biology, 111 Koshland Hall, University of
California, Berkeley, CA 94720. Phone: (510) 642-3906. Fax: (510)
642-9017. E-mail: andyoj{at}uclink4.berkeley.edu.
Journal of Virology, October 2001, p. 9393-9406, Vol. 75, No. 19
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.19.9393-9406.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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