Research Paper Volume 12, Issue 7 pp 5751—5763

Molecular anatomy of the subcellular localization and nuclear import mechanism of herpes simplex virus 1 UL6

Mingsheng Cai 1, *, , Xiaowen Ou 1, *, , Yiwen Li 1, *, , Xingmei Zou 1, , Zuo Xu 1, , Yuanfang Wang 1, , Hao Peng 1, , Yangxi Deng 1, , Yingjie Guo 1, , Manjiao Lu 1, , Weidong Gan 1, , Tao Peng 2, 3, , Meili Li 1, ,

  • 1 Guangdong Provincial Key Laboratory of Allergy and Clinical Immunology, Second Affiliated Hospital of Guangzhou Medical University, Sino-French Hoffmann Institute, School of Basic Medical Science, Guangzhou Medical University, The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People’s Hospital, Guangzhou 510260, Guangdong, China
  • 2 State Key Laboratory of Respiratory Diseases, Sino-French Hoffmann Institute, School of Basic Medical Science, Guangzhou Medical University, Panyu, Guangzhou 511436, Guangdong, China
  • 3 South China Vaccine Corporation Limited, Guangzhou Science Park, Guangzhou 510663, Guangdong, China
* Equal contribution and Co-first authors

received: July 12, 2019 ; accepted: February 8, 2020 ; published: April 1, 2020 ;

https://doi.org/10.18632/aging.102965
How to Cite

Copyright © 2020 Cai et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

As an indispensable structure protein, the herpes simplex virus 1 (HSV-1) UL6 has been described to exert numerous roles in viral proliferation. However, its exact subcellular localization and subcellular transport mechanism is not well known. In the present study, by utilizing confocal fluorescent microscopy, UL6 was shown to mainly locate in the nucleus in enhanced yellow fluorescent protein or Flag tag fused expression plasmid-transfected cells or HSV-1-infected cells, whereas its predicted nuclear localization signal was nonfunctional. In addition, by exploiting dominant negative mutant and inhibitor of different nuclear import receptors, as well as co-immunoprecipitation and RNA interference assays, UL6 was established to interact with importin α1, importin α7 and transportin-1 to mediate its nuclear translocation under the help of Ran-mediated GTP hydrolysis. Accordingly, these results will advance the knowledge of UL6-mediated biological significances in HSV-1 infection cycle.

Abbreviations

aa: amino acids; Co-IP: Co-immunoprecipitation; DN: Dominant negative; EYFP: enhanced yellow fluorescent protein; FITC: fluorescein isothiocyanate; HSV-1: Herpes simplex virus 1; IB: immunoblotting; IFA: Immunofluorescence assay; mAb: monoclonal antibody; NLS: nuclear localization signal; NPC: nuclear pore complex; pAb: polyclonal antibody; shRNA: short hairpin RNA.