Fluorescent tagging of herpes simplex virus tegument protein VP13/14 in virus infection

被引:48
作者
Donnelly, M [1 ]
Elliott, G [1 ]
机构
[1] Marie Curie Res Inst, Viral Assembly Grp, Surrey RH8 0TL, England
关键词
D O I
10.1128/JVI.75.6.2575-2583.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The cellular site of herpesvirus tegument assembly has yet to be defined. We have previously used a recombinant herpes simplex virus type 1 expressing a green fluorescent protein (GFP)-tagged tegument protein, namely VP22, to show that VP22 is localized exclusively to the cytoplasm during infection. Here we have constructed a similar virus expressing another fluorescent tegument protein, YFP-VP13/14, and have visualized the intracellular localization of this second tegument protein in live infected cells, In contrast to VP22, VP13/14 is targeted predominantly to the nuclei of infected cells at both early and late times in infection, More specifically, YFP-13/14 localizes initially to the nuclear replication compartments and then progresses into intense punctate domains that appear at around 12 h postinfection, At even later times this intranuclear punctate fluorescence is gradually replaced by perinuclear micropunctate and membranous fluorescence. While the vast majority of YFP-13/14 seems to be targeted to the nucleus, a minor subpopulation also appears in a vesicular pattern in the cytoplasm that closely resembles the pattern previously observed for GFP-22, Moreover, at late times weak fluorescence appears at the cell periphery and in extracellular virus particles, confirming that YFP13/14 is assembled into virions. This predominantly nuclear targeting of YFP-13/14 together with the cytoplasmic targeting of VP22 may imply that there are multiple sites of tegument protein incorporation along the virus maturation pathway. Thus, our YFP-13/14-expressing virus has revealed the complexity of the intracellular targeting of VP13/14 and provides a novel insight into the mechanism Of tegument, and hence virus, assembly.
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页码:2575 / 2583
页数:9
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