Identification of molecular determinants for the nuclear import of pseudorabies virus UL31

Li, Meili; Jiang, Siwen; Mo, Chuncong; Zeng, Zhan-Cheng; Li, Xiaowei; Chen, Chunke; Yang, Yanjia; Wang, Jinlin; Huang, Jinlu; Chen, Daixiong; Peng, Tao; Cai, Mingsheng

doi:10.1016/j.abb.2015.09.024

Cited by 22 publications

(19 citation statements)

References 54 publications

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“…Co-immunoprecipitation (Co-IP) and immunoblot (IB) were performed as described previously [19,20,23,34,38,39]. HEK293T cells were co-transfected with 10 μg of each of the indicated expression plasmids carrying FLAG or EYFP tag.…”

Section: Co-immunoprecipitation and Immunoblotmentioning

confidence: 99%

Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2

Chen

Zou

et al. 2018

Cell Physiol Biochem

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Background/Aims: Epstein-Barr virus (EBV) BFLF2, the homologue of herpes simplex virus 1 (HSV-1) UL31, is crucial for the efficient viral DNA packaging and primary egress across the nuclear membrane. However, we still do not know its subcellular transport mechanisms. Methods: Interspecies heterokaryon assays were utilized to detect the nucleocytoplasmic shuttling of BFLF2, and mutation analysis, plasmid transfection and fluorescence microscopy assays were performed to identify the functional nuclear localization sequence (NLS) and nuclear export sequence (NES) of BFLF2 in live cells. Furthermore, the nuclear import and export of BFLF2 were assessed by confocal microscopy, co-immunoprecipitation and immunoblot assays. Results: BFLF2 was confirmed to shuttle between the nucleus and cytoplasm. Two predicted NESs were shown to be nonfunctional, yet we proved that the nuclear export of BFLF2 was mediated through transporter associated with antigen processing (TAP), but not chromosomal region maintenance 1 (CRM1) dependent pathway. Furthermore, one functional NLS, ²²RRLMHPHHRNYTASKASAH⁴⁰, was identified, and the aa22-23, aa22-25, aa28-30 and aa37-40 had an important role in the nuclear localization of BFLF2. Besides, the nuclear import of BFLF2 was demonstrated through Ran-, importin α7-, importin β1- and transportin-1-dependent mechanism that does not require importin α1, α3 and α5. Conclusion: These works are of significance for the further study of the functions of BFLF2 during EBV infection, as well as for further insights into the design of new antiviral drug target and vaccine development against EBV.

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Section: Co-immunoprecipitation and Immunoblotmentioning

confidence: 99%

Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2

Chen

Zou

et al. 2018

Cell Physiol Biochem

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“…To eventually determine the minimum NLS region of aa1-43, pEYFP-BLLF2(6-31) and pEYFP-BLLF2 (16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31) were constructed ( Fig. 3A) and transfected into COS-7 cells.…”

Section: Identi Cation Of the Functional Nls In Bllf2mentioning

confidence: 99%

“…6G). As positive controls [25,31], EYFP-BFLF2 and PRV UL31-EYFP could be Co-IPed with anti-Flag mAb when HEK293T cells were co-transfected with plasmids expressing EYFP-BFLF2/3×Flag-importin β1 (Fig. 6H) or PRV UL31-EYFP/3×Flag-importin β1 (Fig.…”

Section: Bllf2 Binds To Importin β1mentioning

confidence: 99%

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Nuclear Import of Epstein-Barr virus BLLF2 is Mediated by an Importin β1-Dependent Mechanism

Guo

Deng

et al. 2020

Preprint

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Background: Epstein-Barr virus (EBV), the pathogen of several human malignancies, encodes many proteins that require to be transported into the nucleus for viral DNA reproduction and nucleocapsids assembly in the lytic replication cycle. A nuclear membrane phosphoprotein encoded by EBV BLLF2, is believed to associate with viral DNA packaging and primary egress across the nuclear membrane. Results: Here, fluorescence microscope, mutation analysis, interspecies heterokaryon assays, co-immunoprecipitation assays and western blot were performed to explore the nuclear import mechanism of BLLF2. As results, BLLF2 was shown to be a nucleocytoplasmic shuttling protein, which was mediated neither by chromosomal region maintenance 1 (CRM1)- nor transporter associated with antigen processing (TAP)-dependent pathway. Yet, two functional nuclear localization signals (NLSs) of BLLF2, NLS1 (16KRQALETVPHPQNRGR31) and NLS2 (48PPVAKRRR58), were identified, whereas the predicted NES was nonfunctional. Finally, BLLF2 was proved to transport into the nucleus via Ran-dependent and importin β1-dependent pathway. Conclusions: This mechanism may contribute to a more extensive insight of the assembly and synthesis of EB virions in the nucleus, thus affording a new direction for the treatment of viruses.

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“…Transfection and fluorescence microscopy procedures were carried out as reported in our previous researches (6)(7)(8)(13)(14)(15). For duplicate the assay, each transfection was manipulated at least three times.…”

Section: Plasmid Transfection and Fluorescence Microscopymentioning

confidence: 99%

Effects of Heat Shock and 2, 4-D Treatment on Morphological and Physiological Characteristics of Microspores and Microspore-Derived Doubled Haploid Plants in Brassica napus L.

et al. 2015

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Identification of molecular determinants for the nuclear import of pseudorabies virus UL31

Cited by 22 publications

References 54 publications

Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2

Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2

Nuclear Import of Epstein-Barr virus BLLF2 is Mediated by an Importin β1-Dependent Mechanism

Effects of Heat Shock and 2, 4-D Treatment on Morphological and Physiological Characteristics of Microspores and Microspore-Derived Doubled Haploid Plants in Brassica napus L.

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