Interactions and nuclear import of the N and P proteins of sonchus yellow net virus, a plant nucleorhabdovirus

被引:63
作者
Goodin, MM [1 ]
Austin, J [1 ]
Tobias, R [1 ]
Fujita, M [1 ]
Morales, C [1 ]
Jackson, AO [1 ]
机构
[1] Univ Calif Berkeley, Dept Plant & Microbial Biol, Berkeley, CA 94720 USA
关键词
D O I
10.1128/JVI.75.19.9393-9406.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:9393 / 9406
页数:14
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