The Role of the Equine Herpesvirus Type 1 (EHV-1) US3-Encoded Protein Kinase in Actin Reorganization and Nuclear Egress

Proft, Alexandra; Spiesschaert, Bart; Izume, Satoko; Taferner, Selina; Lehmann, Maik J.; Azab, Walid

doi:10.3390/v8100275

Cited by 12 publications

(18 citation statements)

References 47 publications

(74 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…6f). The vesicle-like structure observed in the US3-deleted mutant-infected cells is consistent with previous studies in other herpesviruses showing that US3 deletion caused primary enveloped virion accumulation 28,[37][38][39][40] . When US3 was repaired in the revertant virus, virion accumulation in the nucleus was reduced, and the vesicle-like structure disappeared ( Fig.…”

Section: Effect Of the Dpv Us3 Protein On Viral Growth Kineticssupporting

confidence: 91%

The Pivotal Roles of US3 Protein in Cell-to-Cell Spread and Virion Nuclear Egress of Duck Plague Virus

Deng

Wang

Cheng

et al. 2020

Sci Rep

View full text Add to dashboard Cite

The duck plague virus (DPV) US3 protein, a homolog of the herpes simplex virus-1 (HSV-1) US3 protein that is reported to be critical for viral replication, has been minimally studied. Therefore, to investigate the function of the DPV US3 protein, we used scarless Red recombination technology based on an infectious bacterial artificial chromosome (BAC) containing the DPV Chinese virulent strain (CHv) genome and successfully constructed and rescued a US3-deleted mutant and the corresponding revertant virus (BAC-CHv-ΔUS3 and BAC-CHv-ΔUS3R, respectively). For viral growth characteristics, compared to the parental and revertant viruses, the US3-deleted mutant showed an approximately 100-fold reduction in viral titers but no significant reduction in genome copies, indicating that the US3-deleted mutant exhibited decreased viral replication but not decreased viral DNA generation. In addition, the US3-deleted mutant formed viral plaques that were 33% smaller on average than those formed by the parental and revertant viruses, demonstrating that US3 protein affected the viral cell-tocell spread of DPV. Finally, the results of electron microscopy showed that the deletion of US3 resulted in a large number of virions accumulating in the nucleus and perinuclear space, thus blocking virion nuclear egress. In this study, we found that the DPV US3 protein played pivotal roles in viral replication by promoting viral cell-to-cell spread and virion nuclear egress, which may provide some references for research on the function of the DPV US3 protein.Duck plague, also known as duck viral enteritis (DVE), is an acute, febrile, septic disease that appears in waterfowl and is caused by duck plague virus (DPV). Duck plague spreads rapidly with high morbidity and mortality, resulting in significant economic losses in the avian industry. DPV belongs to the Alphaherpesvirinae subfamily and contains a double-stranded helical DNA consisting of a unique long (UL) region, a unique short (US) region, a unique short internal repeat (IRS) region and a unique short terminal repeat (TRS) region, thus forming the UL-IRS-US-TRS structure of the viral genome 1-3 .The characteristics of some genes of DPV have been reported. Herpesvirus genes are classified into immediate-early (IE), early (E) and late (L) genes according to their order of gene expression and thereby play different roles in viral replication. The kinetic classes of many DPV genes have been identified. DPV UL54, an IE gene, can shuttle between the nucleus and cytoplasm to regulate viral replication, and the recombinant UL54-deleted virus produced smaller viral plaque sizes and lower viral genome copies than the parental virus 4-7 . DPV UL13 is an E gene and localizes to the nucleus and cytoplasm, and knocking out UL13 impaired viral replication 8 . Most DPV genes, including US2 9 , US5 10 , US10 11 , UL16 12 , UL35 13 , UL41 14 , UL53 15 , and UL55 16 , are classified as L genes. To date, only a few proteins encoded by DPV apart from UL54 and UL13 have been studied in mutant viruses. Both ...

show abstract

Section: Effect Of the Dpv Us3 Protein On Viral Growth Kineticssupporting

confidence: 91%

The Pivotal Roles of US3 Protein in Cell-to-Cell Spread and Virion Nuclear Egress of Duck Plague Virus

Deng

Wang

Cheng

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…The pattern of NEC localization in ΔUs3 infected cells appeared distinct from that observed in Δ21 infected cells, with a more uniform, regularly spaced arrangement of puncta along the nuclear rim. A similar pattern of NEC localization has been observed in cells infected with Us3 deficient strains of several alphaherpesviruses (3, 15–17). The differences in NEC localization observed between Δ16, Δ21 and ΔUs3 HSV-2 strains suggests that pUL16, pUL21 and pUs3 play roles that are distinct from one another in the nuclear egress of HSV-2.…”

Section: Resultssupporting

confidence: 77%

“…Ultrastructural examination of UL21 mutant strains revealed a number of nuclear envelope perturbations including extravagations of the nuclear envelope into the cytoplasm as well as invaginations of the nuclear envelope and the INM into the nucleoplasm (Fig 6). When INM invaginations were observed, PEVs were often seen accumulating in these structures that were reminiscent of INM herniations seen in cells infected with Us3 mutants derived from PRV (17), EHV-1 (15), MDV (16), HSV-1 (3) and HSV-2 (Fig 11A). Knockdown of pUL31 in cells infected with Δ21 prevented these nuclear envelope perturbations indicating that pUL31 activity was required to drive the formation of these structures and further suggesting that NEC activity is dysregulated in the absence of pUL21.…”

Section: Discussionmentioning

confidence: 97%

“…The interpretation of this observation was that PEV accumulations result as a consequence of the failure of PEVs to de-envelop at the ONM. This interpretation has been widely utilized to explain the phenotypes of Us3 mutant strains derived from other herpesviruses including HSV-1, EHV-1 and MDV (3, 15, 16). The data presented here confound this interpretation.…”

Section: Discussionmentioning

confidence: 99%

“…One protein that is capable of regulating NEC activity is the viral serine/threonine kinase pUs3. In cells infected with pUs3 mutants of pseudorabies virus (PRV), HSV-1, equine herpes virus 1 (EHV-1) and Marek’s disease virus (MDV) PEVs accumulate in herniations of the INM (3, 15–17). HSV-1 pUs3 phosphorylates pUL31 at a number of serine residues in its N-terminus and it has been proposed that these modifications are required for efficient de-envelopment of PEVs at the ONM, because mutation of these pUL31 serine residues to alanine recapitulates the pUs3 mutant phenotype (18).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Differentiating the Roles of UL16, UL21 and Us3 in the Nuclear Egress of Herpes Simplex Virus Capsids

Gao

Finnen

Sherry

et al. 2020

Preprint

View full text Add to dashboard Cite

AbstractPrevious studies from our laboratory established that pUL16 and pUL21 are required for efficient nuclear egress of herpes simplex type 2 (HSV-2) capsids. To better understand the role of these proteins in nuclear egress, we wished to establish whether nuclear egress complex (NEC) localization and/or function was altered in the absence of either pUL16 or pUL21. We used antiserum raised against HSV-2 NEC components pUL31 and pUL34 to examine NEC localization by immunofluorescence microscopy. NEC localization in cells infected with pUL16 deficient viruses was indistinguishable from that observed in cells infected with wild type viruses. By contrast, NEC localization was found to be aberrant in cells infected with pUL21 deficient virus and, instead, showed some similarity to the aberrant NEC localization pattern observed in cells infected with pUs3 deficient virus. These results indicated that pUL16 plays a role in nuclear egress that is distinct from that of pUL21 and pUs3. Higher resolution examination of nuclear envelope ultrastructure in cells infected with pUL21 deficient viruses by transmission electron microscopy showed different types of nuclear envelope perturbations, including some that were not observed in cells infected with pUs3 deficient virus. The formation of the nuclear envelope perturbations observed in pUL21 deficient virus infections was found to be dependent on a functional NEC, revealing a novel role for pUL21 in regulating NEC activity. The results of comparisons of nuclear envelope ultrastructure in cells infected with viruses lacking pUs3, pUL16 or both pUs3 and pUL16 were consistent with a role for pUL16 upstream of primary capsid envelopment and shed new light on how pUs3 functions in nuclear egress.Author summaryThe membrane deformation activity of the herpesvirus nuclear egress complex (NEC), allows viral capsids to transit from their site of assembly in the nucleus through both nuclear membranes into the cytoplasm. The timing, extent and directionality of NEC activity must be precisely controlled during viral infection, yet our knowledge of how NEC activity is controlled is incomplete. To determine how pUL16 and pUL21, two viral proteins required for nuclear egress of herpes simplex virus type 2 (HSV-2) capsids, function to promote nuclear egress, we examined how the lack of each protein impacted NEC localization. These analyses revealed a function of pUL16 in nuclear egress that is distinct from that of pUL21, uncovered a novel role for pUL21 in regulating NEC activity and shed new light on how a viral kinase, pUs3, regulates nuclear egress. Nuclear egress of viral capsids is a common feature of the replicative cycle of all herpesviruses. A complete understanding of all aspects of nuclear egress, including how viral NEC activity is controlled, may yield strategies to disrupt this process that could be applied to the development of herpes-specific antiviral drugs.

show abstract

Equid Herpesvirus-1 Exploits the Extracellular Matrix of Mononuclear Cells to Ensure Transport to Target Cells

et al. 2020

Self Cite

View full text Add to dashboard Cite

Mononuclear cells are the first line of defense against microbial infection. Yet, several viruses have evolved different mechanisms to overcome host defenses to ensure their spread. Here, we show unique mechanisms of how equid herpesvirus-1 manipulates peripheral blood mononuclear cells (PBMC) to travel further in the body. (i) “PBMC-hitching”: at the initial contact, herpesviruses lurk in the extracellular matrix (ECM) of PBMC without entering the cells. The virus exploits the components of the ECM to bind, transport and then egress to infect other cells. (ii) “Intracellular delivery”: transendothelial migration is a physiological mechanism where mononuclear cells can transmigrate through the endothelial cells. The virus was intangible and probably did not interfere with such a mechanism where the infected PBMC can probably deliver the virus inside the endothelium. (iii) “Classical-fusion”: this process is well mastered by herpesviruses due to a set of envelope glycoproteins that facilitate cell-cell fusion and virus spread.

show abstract

The Role of the Equine Herpesvirus Type 1 (EHV-1) US3-Encoded Protein Kinase in Actin Reorganization and Nuclear Egress

Cited by 12 publications

References 47 publications

The Pivotal Roles of US3 Protein in Cell-to-Cell Spread and Virion Nuclear Egress of Duck Plague Virus

The Pivotal Roles of US3 Protein in Cell-to-Cell Spread and Virion Nuclear Egress of Duck Plague Virus

Differentiating the Roles of UL16, UL21 and Us3 in the Nuclear Egress of Herpes Simplex Virus Capsids

Equid Herpesvirus-1 Exploits the Extracellular Matrix of Mononuclear Cells to Ensure Transport to Target Cells

Contact Info

Product

Resources

About