Herpes simplex virus 1 recombinant virions exhibiting the amino terminal fragment of urokinase-type plasminogen activator can enter cells via the cognate receptor

ABSTRACT Earlier studies have shown that the herpes simplex virus (HSV) US3 encodes two transcriptional units directing the synthesis of the US3 (residues 1 to 481) and US3.5 (residues 77 to 481) protein kinases. Both kinases phosphorylate histone deacetylase 1 (HDAC1) and HDAC2 and enable the expression of genes cotransduced into U2OS cells by recombinant baculoviruses, an activity designated the “helper function.” The two kinases differ with respect to antiapoptotic activity. In the studies reported here, we made a series of FLAG-tagged amino- and carboxyl-terminal truncations of US3 and these were tested for antiapoptotic activity, phosphorylation of HDAC1, and the helper function. We report the following. (i) HDAC1 phosphorylation and the helper function were expressed in cells transduced by the truncation encoding residues 182 to 481 but not in cells transduced by the truncation encoding residues 189 to 481 or the amino-terminal polypeptides encompassing the first 188 amino acids. (ii) The self-posttranslational modification requires residues 164 to 481. (iii) The antiapoptotic activity requires both the amino-terminal and the carboxyl-terminal domains, of which the truncated protein containing residues 1 to 163 and that containing residues 164 to 481, respectively, were the smallest fragments tested to be effective. The two domains need not be on the same molecule, but they must overlap. The smallest overlapping pair tested was the fragment containing residues 1 to 181 and that containing residues 164 to 481. Consistent with the hypothesis that the effective overlapping truncations form a heteromultimeric structure, antibody to FLAG coprecipitated untagged US3 from lysates of cells cotransduced with FLAG-tagged, truncated US3 constructs. Although US3 has been reported to be a monomeric enzyme, the results indicate that it can form enzymatically active multimeric structures.

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Functional Comparison of Herpes Simplex Virus 1 (HSV-1) and HSV-2 ICP27 Homologs Reveals a Role for ICP27 in Virion Release

Journal of Virology ◽

10.1128/jvi.02994-14 ◽

2014 ◽

Vol 89 (5) ◽

pp. 2892-2905 ◽

Cited By ~ 10

Author(s):

Donglim Park ◽

Joseph Lalli ◽

Lenka Sedlackova-Slavikova ◽

Stephen A. Rice

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Infected Cell ◽

Regulatory Protein ◽

Late Gene Expression ◽

Herpes Simplex Virus 1 ◽

Amino Terminal ◽

Virion Release ◽

Simplex Virus ◽

Hsv 1

ABSTRACTNumerous studies have focused on the regulatory functions of ICP27, an immediate-early (IE) protein of herpes simplex virus 1 (HSV-1). However, its homolog in HSV-2, termed ICP27t2, has been little studied. Here, we used two different approaches to functionally compare ICP27t2 and ICP27. In transfection-based assays, ICP27t2 closely resembled ICP27 in its capacity to enhance HSV-1 late gene expression, suppress the splicing of a viral intron, and complement the growth of an HSV-1 ICP27 null mutant. To study ICP27t2 in the context of viral infection, we engineered K2F1, an HSV-1 mutant that encodes ICP27t2 in place of ICP27. In Vero cells, K2F1 replicated with wild-type (WT) kinetics and yields, expressed delayed-early and late proteins normally, and was fully capable of activating several cellular signal transduction pathways that are ICP27 dependent. Thus, we conclude that ICP27t2 and ICP27 are functionally very similar and that ICP27t2 can mediate all ICP27 activities that are required for HSV-1 replication in cell culture. Surprisingly, however, we found that K2F1 forms plaques that are morphologically different from those of WT HSV-1. Investigation of this trait demonstrated that it results from the decreased release of progeny virions into the culture medium. This appears to be due to a reduction in the detachment of K2F1 progeny from the extracellular surface of the infected cell. We identified two HSV-1 ICP27 amino-terminal deletion mutants with a similar release defect. Together, these results demonstrate that ICP27 plays a heretofore-unappreciated role in modulating the efficiency of progeny virion release.IMPORTANCEICP27 is an essential, multifunctional regulatory protein that has a number of critical roles in the HSV-1 life cycle. Although ICP27 homologs are encoded by all known members of theHerpesviridae, previous work with several of these homologs has shown that they cannot substitute for ICP27 in the context of HSV-1-infected cells. Here, we identify ICP27t2 as the first homolog that can efficiently replace ICP27 in HSV-1 infection. Unexpectedly, our results also reveal that the sequence of the ICP27 gene can affect the release of HSV-1 progeny virions from the infected cell. Thus, our comparative study has revealed a novel function for ICP27 in the regulation of virus release.

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Amino-Terminal Fragment of Urokinase-Type Plasminogen Activator Inhibits HIV-1 Replication

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.2001.4965 ◽

2001 ◽

Vol 284 (2) ◽

pp. 346-351 ◽

Cited By ~ 21

Author(s):

Manabu Wada ◽

Naoko A. Wada ◽

Hiroyuki Shirono ◽

Katsumi Taniguchi ◽

Hideaki Tsuchie ◽

...

Keyword(s):

Plasminogen Activator ◽

Amino Terminal ◽

Terminal Fragment ◽

Type Plasminogen Activator ◽

Urokinase Type Plasminogen Activator ◽

Hiv 1 ◽

Amino Terminal Fragment

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Characterization of the triplet repeats in the central domain of the γ 134·5 protein of herpes simplex virus 1

Journal of General Virology ◽

10.1099/vir.0.81033-0 ◽

2005 ◽

Vol 86 (9) ◽

pp. 2411-2419 ◽

Cited By ~ 5

Author(s):

Xianghong Jing ◽

Bin He

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Triplet Repeats ◽

Herpes Simplex Virus 1 ◽

Central Domain ◽

Amino Terminal ◽

Terminal Domain ◽

Different Strains ◽

Simplex Virus ◽

Hsv 1

The γ 134·5 protein of herpes simplex virus 1 (HSV-1) consists of an amino-terminal domain, a central domain with triplet repeats (Ala–Thr–Pro) and a carboxyl-terminal domain. The triplet repeats are a unique feature of the γ 134·5 protein encoded by HSV-1, but the number of repeats varies among different strains. Notably, the central domain containing the triplet repeats is implicated in neuroinvasion. In this report, it has been shown that partial or full deletion of triplet repeats, i.e. from ten to either three or zero, in the γ 134·5 protein has no effect on the virus response to interferon. The triplet deletion mutants replicate efficiently in CV-1 and mouse 10T1/2 cells. However, in mouse 3T6 cells, these mutants grow with delayed growth kinetics. This decrease in growth, compared with wild-type HSV-1(F), does not result from failure of the virus to suppress the RNA-dependent protein kinase response, but rather from a delay in virus release or egress. Accordingly, these mutant viruses are predominantly present within infected cells. These results indicate that deletions in the central domain of the γ 134·5 protein impair virus egress, but not virus response to interferon.

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Role of the arginine cluster in the disordered domain of Herpes Simplex Virus 1 UL34 for the recruitment of ESCRT-III for viral primary envelopment

Journal of Virology ◽

10.1128/jvi.01704-21 ◽

2021 ◽

Author(s):

Jun Arii ◽

Kosuke Takeshima ◽

Yuhei Maruzuru ◽

Naoto Koyanagi ◽

Yoshitaka Nakayama ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Nuclear Export ◽

Herpes Simplex Virus 1 ◽

Amino Terminal ◽

Arginine Residues ◽

Infected Cells ◽

Simplex Virus ◽

Hsv 1

During the nuclear export of nascent nucleocapsids of herpesviruses, the nucleocapsids bud through the inner nuclear membrane (INM) by acquiring the INM as a primary envelope (primary envelopment). We recently reported that herpes simplex virus 1 (HSV-1) nuclear egress complex (NEC), which consists of UL34 and UL31, interacts with an ESCRT-III adaptor ALIX and recruits ESCRT-III machinery to the INM for efficient primary envelopment. In this study, we identified a cluster of six arginine residues in the disordered domain of UL34 as a minimal region required for the interaction with ALIX as well as the recruitment of ALIX and an ESCRT-III protein CHMP4B to the INM in HSV-1-infected cells. Mutations in the arginine cluster exhibited phenotypes similar to those with ESCRT-III inhibition reported previously, including the mis-localization of NEC, induction of membranous invagination structures containing enveloped virions, aberrant accumulation of enveloped virions in the invaginations and perinuclear space, and reduction of viral replication. We also showed that the effect of the arginine cluster in UL34 on HSV-1 replication was dependent primarily on ALIX. These results indicated that the arginine cluster in the disordered domain of UL34 was required for the interaction with ALIX and the recruitment of ESCRT-III machinery to the INM to promote primary envelopment. IMPORTANCE Herpesvirus UL34 homologs contain conserved amino-terminal domains that mediate vesicle formation through interactions with UL31 homologs during primary envelopment. UL34 homologs also comprise other domains adjacent to their membrane-anchoring regions, which differ in length, are variable in herpesviruses and do not form distinguished secondary structures. However, the role of these disordered domains in infected cells remains to be elucidated. In this study, we present data suggesting that the arginine cluster in the disordered domain of HSV-1 UL34 mediates the interaction with ALIX, thereby leading to the recruitment of ESCRT-III machinery to the INM for efficient primary envelopment. This is the first study to report the role of the disordered domain of a UL34 homolog in herpesvirus infections.

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Herpes Simplex Virus 1 γ134.5 Protein Inhibits STING Activation That Restricts Viral Replication

Journal of Virology ◽

10.1128/jvi.01015-18 ◽

2018 ◽

Vol 92 (20) ◽

Cited By ~ 20

Author(s):

Shuang Pan ◽

Xing Liu ◽

Yijie Ma ◽

Youjia Cao ◽

Bin He

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virulence Factor ◽

Dna Sensing ◽

Viral Inhibition ◽

Herpes Simplex Virus 1 ◽

Amino Terminal ◽

Viral Virulence ◽

Simplex Virus ◽

Hsv 1

ABSTRACTThe γ134.5 gene of herpes simplex virus 1 (HSV-1) encodes a virulence factor that promotes viral pathogenesis. Although it perturbs TANK-binding kinase 1 (TBK1) in the complex network of innate immune pathways, the underlying mechanism is obscure. Here we report that HSV-1 γ134.5 targets stimulator of interferon genes (STING) in the intracellular DNA recognition pathway that regulates TBK1 activation. In virus-infected cells the γ134.5 protein associates with and inactivates STING, which leads to downregulation of interferon regulatory factor 3 (IRF3) and IFN responses. Importantly, HSV-1 γ134.5 disrupts translocation of STING from the endoplasmic reticulum to Golgi apparatus, a process necessary to prime cellular immunity. Deletion of γ134.5 or its amino-terminal domain from HSV-1 abolishes the observed inhibitory activities. Consistently, an HSV mutant that lacks functional γ134.5 replicated less efficiently in STING+/+than in STING−/−mouse embryonic fibroblasts. Moreover, reconstituted expression of human STING in the STING−/−cells activated IRF3 and reduced viral growth. These results suggest that control of the DNA sensing pathway by γ134.5 is advantageous to HSV infection.IMPORTANCEViral inhibition of innate immunity contributes to herpes simplex virus pathogenesis. Although this complex process involves multiple factors, the underlying events remain unclear. We demonstrate that an HSV virulence factor γ134.5 precludes the activation of STING, a central adaptor in the intracellular DNA sensing pathway. Upon HSV infection, this viral protein engages with and inactivates STING. Consequently, it compromises host immunity and facilitates HSV replication. These observations uncover an HSV mechanism that is likely to mediate viral virulence.

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cdc2 Cyclin-Dependent Kinase Binds and Phosphorylates Herpes Simplex Virus 1 UL42 DNA Synthesis Processivity Factor

Journal of Virology ◽

10.1128/jvi.75.21.10326-10333.2001 ◽

2001 ◽

Vol 75 (21) ◽

pp. 10326-10333 ◽

Cited By ~ 30

Author(s):

Sunil J. Advani ◽

Ralph R. Weichselbaum ◽

Bernard Roizman

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Kinase Activity ◽

Dominant Negative ◽

Cdc2 Kinase ◽

Herpes Simplex Virus 1 ◽

Amino Terminal ◽

Processivity Factor ◽

Carboxyl Terminal ◽

Simplex Virus

ABSTRACT Earlier studies have shown that cdc2 kinase is activated during herpes simplex virus 1 infection and that its activity is enhanced late in infection even though the levels of cyclin A and B are decreased below levels of detection. Furthermore, activation of cdc2 requires the presence of infected cell protein no. 22 and the UL13 protein kinase, the same gene products required for optimal expression of a subset of late genes exemplified by US11, UL38, and UL41. The possibility that the activation of cdc2 and expression of this subset may be connected emerged from the observation that dominant negative cdc2 specifically blocked the expression of US11 protein in cells infected and expressing dominant negative cdc2. Here we report that in the course of searching for a putative cognate partner for cdc2 that may have replaced cyclins A and B, we noted that the DNA polymerase processivity factor encoded by the UL42 gene contains a degenerate cyclin box and has been reported to be structurally related to proliferating cell nuclear antigen, which also binds cdk2. Consistent with this finding, we report that (i) UL42 is able to physically interact with cdc2 at both the amino-terminal and carboxyl-terminal domains, (ii) the carboxyl-terminal domain of UL42 can be phosphorylated by cdc2, (iii) immunoprecipitates obtained with anti UL42 antibody contained a roscovitine-sensitive kinase activity, (iv) kinase activity associated with UL42 could be immunodepleted by antibody to cdc2, and (v) UL42 transfected into cells associates with a nocodazole-enhanced kinase. We conclude that UL42 can associate with cdc2 and that the kinase activity has the characteristic traits of cdc2 kinase.

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