identification of a novel nuclear localization signal of baculovirus late expression factor 11
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- 英文论文题名：identification of a novel nuclear localization signal of baculovirus late expression factor 11
- 论文作者：Jun Zhang ; Zhan-Qi Dong ; Chun-Dong Zhang ; Qian He ; Xue-Mei Chen ; Ming-Ya Cao ; Hai-Qin Li ; Wen-Fu Xiao ; Cheng Lu ; Min-Hui Pan
- 英文论文作者：Jun Zhang ; Zhan-Qi Dong ; Chun-Dong Zhang ; Qian He ; Xue-Mei Chen ; Ming-Ya Cao ; Hai-Qin Li ; Wen-Fu Xiao ; Cheng Lu ; Min-Hui Pan ; State Key Laboratory of Silkworm Genome Biology ; Institute of Sericulture and Systems ; Southwest University ; Department of Biochemistry and Molecular Biology ; Chongqing Medical University ; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry ; Southwest University
- 年：2016
- 作者机构：State Key Laboratory of Silkworm Genome Biology, Institute of Sericulture and Systems, Southwest University;Department of Biochemistry and Molecular Biology, Chongqing Medical University;Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University;
- 英文论文关键词：Bombyx mori ; Nucleopolyhedrovirus ; Nuclear localization signal ; Importin subunit α-3 ; Viral DNA replication site ; Protein–protein interaction
- 会议召开时间：2016-07-01
- 会议录名称：第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会论文集（摘要汇编）
- 语种：英文
- 分类号：S884.5
- 学会代码：ZGCU
- 会议名称：第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会
- 会议地点：中国重庆
- 主办单位：中国蚕学会、国家蚕桑产业技术体系
- 学会名称：中国蚕学会
- 页数：1
- 文件大小：485k
- 原文格式：D
- 会议级别：全国
摘要
The baculovirus late expression factor 11(LEF-11) has been reported to be involved in viral DNA replication and late/very late gene activation. In this study, serial N- and C-terminal truncations of Bombyx mori nucleopolyhedrovirus(BmNPV) LEF-11 protein were fused with Ds Red to investigate the nuclear localization signal by which LEF-11 enters the nucleus. Results show that 72-101 residues at the C-terminus are essential for BmNPV LEF-11 nuclear localization. Sequence alignment of this NLS from multiple LEF-11 homologs revealed high conservation in general. Site-directed mutation analysis showed that five basic residue clusters, namely, K~(75)/R~(76), H~(81), K~(83)/R~(84), R~(87) and K~(100), were critical for the nuclear localization of BmNPV LEF-11. Co-IF analysis shows that LEF-11 binds directly to host importin α-3. Immunofluorescence analysis demonstrated that LEF-11 co-localizes with the immediate-early protein IE-1 at viral DNA replication sites in the nucleus. Further BiFC assays demonstrated the interaction of LEF-11 with LEF-3 and LEF-11 itself in the nucleus. Together, these results reveal a previous unknown mechanism for nuclear translocation of baculovirus LEF-11.
The baculovirus late expression factor 11(LEF-11) has been reported to be involved in viral DNA replication and late/very late gene activation. In this study, serial N- and C-terminal truncations of Bombyx mori nucleopolyhedrovirus(BmNPV) LEF-11 protein were fused with Ds Red to investigate the nuclear localization signal by which LEF-11 enters the nucleus. Results show that 72-101 residues at the C-terminus are essential for BmNPV LEF-11 nuclear localization. Sequence alignment of this NLS from multiple LEF-11 homologs revealed high conservation in general. Site-directed mutation analysis showed that five basic residue clusters, namely, K~(75)/R~(76), H~(81), K~(83)/R~(84), R~(87) and K~(100), were critical for the nuclear localization of BmNPV LEF-11. Co-IF analysis shows that LEF-11 binds directly to host importin α-3. Immunofluorescence analysis demonstrated that LEF-11 co-localizes with the immediate-early protein IE-1 at viral DNA replication sites in the nucleus. Further BiFC assays demonstrated the interaction of LEF-11 with LEF-3 and LEF-11 itself in the nucleus. Together, these results reveal a previous unknown mechanism for nuclear translocation of baculovirus LEF-11.
The baculovirus late expression factor 11(LEF-11) has been reported to be involved in viral DNA replication and late/very late gene activation. In this study, serial N- and C-terminal truncations of Bombyx mori nucleopolyhedrovirus(BmNPV) LEF-11 protein were fused with Ds Red to investigate the nuclear localization signal by which LEF-11 enters the nucleus. Results show that 72-101 residues at the C-terminus are essential for BmNPV LEF-11 nuclear localization. Sequence alignment of this NLS from multiple LEF-11 homologs revealed high conservation in general. Site-directed mutation analysis showed that five basic residue clusters, namely, K~(75)/R~(76), H~(81), K~(83)/R~(84), R~(87) and K~(100), were critical for the nuclear localization of BmNPV LEF-11. Co-IF analysis shows that LEF-11 binds directly to host importin α-3. Immunofluorescence analysis demonstrated that LEF-11 co-localizes with the immediate-early protein IE-1 at viral DNA replication sites in the nucleus. Further BiFC assays demonstrated the interaction of LEF-11 with LEF-3 and LEF-11 itself in the nucleus. Together, these results reveal a previous unknown mechanism for nuclear translocation of baculovirus LEF-11.
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