Comparative proteomics identifies Schlafen 5 (SLFN5) as a herpes simplex virus restriction factor that suppresses viral transcription

Kim, Eui Tae; Dybas, Joseph M.; Kulej, Katarzyna; Reyes, Emigdio D.; Price, Alexander M.; Akhtar, Lisa N.; Orr, A.B.; García, Benjamin A.; Boutell, Chris; Weitzman, Matthew D.

doi:10.1038/s41564-020-00826-3

Cited by 29 publications

(39 citation statements)

References 75 publications

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“…Although the Schlafen family members share many highly conserved sequence regions, their biological roles and enzymatic functions differ. While some family members such as rat Slfn13 and human SLFN11 influence the translation machinery ( 31 , 42 ), others, such as SLFN5, are involved in transcriptional regulation ( 26 , 47 , 50 ). In particular, the molecular mechanism of human SLFN5 in tumor control is not well understood.…”

Section: Discussionmentioning

confidence: 99%

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Structural and biochemical characterization of human Schlafen 5

Metzner¹,

Huber²,

Hopfner³

et al. 2022

Nucleic Acids Research

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The Schlafen family belongs to the interferon-stimulated genes and its members are involved in cell cycle regulation, T cell quiescence, inhibition of viral replication, DNA-repair and tRNA processing. Here, we present the cryo-EM structure of full-length human Schlafen 5 (SLFN5) and the high-resolution crystal structure of the highly conserved N-terminal core domain. We show that the core domain does not resemble an ATPase-like fold and neither binds nor hydrolyzes ATP. SLFN5 binds tRNA as well as single- and double-stranded DNA, suggesting a potential role in transcriptional regulation. Unlike rat Slfn13 or human SLFN11, human SLFN5 did not cleave tRNA. Based on the structure, we identified two residues in proximity to the zinc finger motif that decreased DNA binding when mutated. These results indicate that Schlafen proteins have divergent enzymatic functions and provide a structural platform for future biochemical and genetic studies.

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Section: Discussionmentioning

confidence: 99%

“…It represses HSV-1 transcription by binding to viral DNA and in turn, preventing RNA polymerase II from accessing viral promoters. However, in the presence of the viral E3 ubiquitin ligase ICP0, SLFN5 is ubiquitinated and subject to proteasomal degradation ( 26 ).…”

Section: Introductionmentioning

confidence: 99%

Structural and biochemical characterization of human Schlafen 5

Metzner¹,

Huber²,

Hopfner³

et al. 2022

Nucleic Acids Research

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“…Specifically, ICP0 targets Schlafen 5 (SLFN5) for proteasomal degradation via ubiquitination of SLFN5. Infection with virus lacking ICP0 (HSV-1ΔICP0) SLFN5 binds to viral DNA to repress transcription via limiting RNA polymerase II access to immediate-early viral promoters [ 72 ]. Similarly, TRIM22 has been identified as an intrinsic host cell factor limiting HSV replication via the viral genome’s heterochromatinization.…”

Section: Rational Design Of the Hsv-2 0∆nls Vaccinementioning

confidence: 99%

Rational Design of Live-Attenuated Vaccines against Herpes Simplex Viruses

Stanfield

Kousoulas

Fernández³

et al. 2021

Viruses

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Diseases caused by human herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) affect millions of people worldwide and range from fatal encephalitis in neonates and herpes keratitis to orofacial and genital herpes, among other manifestations. The viruses can be shed efficiently by asymptomatic carriers, causing increased rates of infection. Viral transmission occurs through direct contact of mucosal surfaces followed by initial replication of the incoming virus in skin tissues. Subsequently, the viruses infect sensory neurons in the trigeminal and lumbosacral dorsal root ganglia, where they are primarily maintained in a transcriptionally repressed state termed “latency”, which persists for the lifetime of the host. HSV DNA has also been detected in other sympathetic ganglia. Periodically, latent viruses can reactivate, causing ulcerative and often painful lesions primarily at the site of primary infection and proximal sites. In the United States, recurrent genital herpes alone accounts for more than a billion dollars in direct medical costs per year, while there are much higher costs associated with the socio-economic aspects of diseased patients, such as loss of productivity due to mental anguish. Currently, there are no effective FDA-approved vaccines for either prophylactic or therapeutic treatment of human herpes simplex infections, while several recent clinical trials have failed to achieve their endpoint goals. Historically, live-attenuated vaccines have successfully combated viral diseases, including polio, influenza, measles, and smallpox. Vaccines aimed to protect against the devastation of smallpox led to the most significant achievement in medical history: the eradication of human disease by vaccination. Recently, novel approaches toward developing safe and effective live-attenuated vaccines have demonstrated high efficacy in various preclinical models of herpetic disease. This next generation of live-attenuated vaccines has been tailored to minimize vaccine-associated side effects and promote effective and long-lasting immune responses. The ultimate goal is to prevent or reduce primary infections (prophylactic vaccines) or reduce the frequency and severity of disease associated with reactivation events (therapeutic vaccines). These vaccines’ “rational” design is based on our current understanding of the immunopathogenesis of herpesviral infections that guide the development of vaccines that generate robust and protective immune responses. This review covers recent advances in the development of herpes simplex vaccines and the current state of ongoing clinical trials in pursuit of an effective vaccine against herpes simplex virus infections and associated diseases.

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“…During infection, Pol II is strongly enriched in viral replication compartments (9): large compartments predominately assembled by viral DNA and the viral transcription factor ICP4 which recruit numerous factors for transcription, DNA replication, and virion assembly while excluding host proteins such as histones and gene silencing factors (50). We sought to determine which CTD modifications localized to viral DNA by immunofluorescence.…”

Section: Herpesviral Dna Recruits But Does Not Require All Ctd Modificationsmentioning

confidence: 99%

Herpes simplex virus 1 inhibits phosphorylation of RNA polymerase II CTD serine-7

Dionisi

Grothey

et al. 2021

Preprint

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Transcriptional activity of RNA polymerase II (Pol II) is orchestrated by post-translational modifications of the C-terminal domain (CTD) of the largest Pol II subunit, RPB1. Herpes Simplex Virus type 1 (HSV-1) usurps the cellular transcriptional machinery during lytic infection to efficiently express viral mRNA and shut down host gene expression. The viral immediate-early protein ICP22 interferes with serine 2 phosphorylation (pS2) of the Pol II CTD by targeting CDK9. The functional implications of this are poorly understood. Here, we report that HSV-1 also induces a global loss of serine 7 phosphorylation (pS7). This effect was dependent on the expression of the two viral immediate-early proteins, ICP22 and ICP27. While lytic HSV-1 infection results in efficient Pol II degradation late in infection, we show that pS2/S7 loss precedes the drop in Pol II level. Interestingly, mutation of the RPB1 polyubiquitination site mutation K1268, which prevents proteasomal RPB1 degradation during transcription-coupled DNA repair, displayed loss of pS2/S7 but retained much higher overall RPB1 protein levels even at late times of infection, indicating that this pathway mediates bulk Pol II protein loss late in infection but is not involved in early CTD dysregulation. Using α-amanitin-resistant CTD mutants, we observed differential requirements for Ser2 and Ser7 for production of viral proteins, with Ser2 facilitating viral immediate-early gene expression and Ser7 appearing dispensable. Despite dysregulation of CTD phosphorylation and different requirements for Ser2/7, all CTD modifications tested could be visualized in viral replication compartments by immunofluorescence. These data expand the known means that HSV-1 employs to create pro-viral transcriptional environments at the expense of host responses.

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Comparative proteomics identifies Schlafen 5 (SLFN5) as a herpes simplex virus restriction factor that suppresses viral transcription

Cited by 29 publications

References 75 publications

Structural and biochemical characterization of human Schlafen 5

Structural and biochemical characterization of human Schlafen 5

Rational Design of Live-Attenuated Vaccines against Herpes Simplex Viruses

Herpes simplex virus 1 inhibits phosphorylation of RNA polymerase II CTD serine-7

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