Phosphorylation and nuclear localization of the varicella-zoster virus gene 63 protein.
Phosphorylation and nuclear localization of the varicella-zoster virus gene 63 protein.
复制标题
水痘带状疱疹病毒基因 63 蛋白的磷酸化和核定位。
DOI：
10.1128/jvi.70.1.658-662.1996
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Ruyechan,WT
中科院分区：
文献类型：
--
作者：
Stevenson,D;Xue,M;Hay,J;Ruyechan,WT
由水痘带状疱疹病毒开放阅读框63编码的蛋白质及其羧基末端缺失在大肠杆菌中表达为麦芽糖结合蛋白羧基末端的融合蛋白，或在转染的哺乳动物细胞中独立表达。截短物包含完整的278个氨基酸一级序列的氨基酸1至142（63 Δ N）或1至210（63 Δ K）。重组酪蛋白激酶II在体外磷酸化63 F和63 Δ KF融合蛋白，但不磷酸化63 Δ NF融合蛋白，这意味着磷酸化发生在氨基酸142和210之间。用表达63、63 Δ N或63 Δ K的质粒转染的细胞的35S或32P标记提取物的免疫沉淀也表明原位磷酸化最可能发生在氨基酸142和210之间。这些综合结果表明，酪蛋白激酶II在水痘带状疱疹病毒63蛋白的磷酸化中起着重要作用。转染细胞的间接免疫荧光显示63蛋白的核定位和63 Δ K和63 Δ N的胞质定位，这意味着有效的核定位需要氨基酸210和278之间的序列。
The protein encoded by varicella-zoster virus open reading frame 63 and carboxy-terminal deletions of the same were expressed either as fusion proteins at the carboxy terminus of the maltose-binding protein in Escherichia coli or independently in transfected mammalian cells. The truncations contained amino acids 1 to 142 (63 delta N) or 1 to 210 (63 delta K) of the complete 278-amino-acid primary sequence. Recombinant casein kinase II phosphorylated the 63F and 63 delta KF fusion proteins in vitro but did not phosphorylate the 63 delta NF fusion protein, implying that phosphorylation occurred between amino acids 142 and 210. Immunoprecipitation of 35S- or 32P-labelled extracts of cells transfected with plasmids expressing 63, 63 delta N, or 63 delta K also indicated that in situ phosphorylation most likely occurred between amino acids 142 and 210. These combined results suggest that casein kinase II plays a significant role in the phosphorylation of the varicella-zoster virus 63 protein. Indirect immunofluorescence of transfected cells indicated nuclear localization of the 63 protein and cytoplasmic localization of 63 delta K and 63 delta N, implying a requirement for sequences between amino acids 210 and 278 for efficient nuclear localization.