Proteomic Screening of Human Targets of Viral microRNAs Reveals Functions Associated with Immune Evasion and Angiogenesis

Gallaher, Amelia M.; Das, Sudipto; Xiao, Zhen; Andrésson, Þorkell; Kieffer-Kwon, Philippe; Happel, Christine; Ziegelbauer, Joseph M.

doi:10.1371/journal.ppat.1003584

Cited by 52 publications

(48 citation statements)

References 56 publications

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“…1A). We note that previous data demonstrated miRNA mimics are not overrepresented in the RNA-induced silencing complex, as determined by argonaute immunoprecipitation experiments (23,24). The reduced expression of GADD45B suggested that this gene was directly targeted for repression by KSHV miRNAs.…”

Section: Resultssupporting

confidence: 52%

“…We utilized our previously published data sets that investigated changes in human gene expression in response to KSHV infection or in separate assays with cells transfected with KSHV miRNA mimics. We focused on mRNA expression changes after KSHV infection (22) or after transfection of a pool of KSHV miRNAs (23) and found that the growth arrest and DNA-damage-inducible 45 beta (GADD45B) gene was the 9th most repressed gene out of 13,795 microarray probes when combining these two data sets (Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

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Kaposi's Sarcoma-Associated Herpesvirus MicroRNAs Target GADD45B To Protect Infected Cells from Cell Cycle Arrest and Apoptosis

Liu

Happel

Ziegelbauer

2017

J Virol

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Kaposi's sarcoma is one of the most common malignancies in HIVinfected individuals. The responsible agent, Kaposi's sarcoma-associated herpesvirus (KSHV; HHV8), expresses multiple microRNAs (miRNAs), but the targets and functions of these miRNAs are not completely understood. After infection in primary endothelial cells with KSHV, growth arrest DNA damage-inducible gene 45 beta (GADD45B) is one of the most repressed genes using genomic expression profiling. GADD45B was also repressed in mRNA expression profiling experiments when KSHV miRNAs were introduced to uninfected cells. We hypothesized that KSHV miRNAs target human GADD45B to protect cells from consequences of DNA damage, which can be triggered by viral infection. Expression of GADD45B protein is induced by the p53 activator, Nutlin-3, and KSHV miRNA-K9 inhibits this induction. In addition, Nutlin-3 increased apoptosis and cell cycle arrest based on flow cytometry assays. KSHV miR-K9 protected primary endothelial cells from apoptosis and cell cycle arrest following Nutlin-3 treatment. Similar protective phenotypes were seen for targeting GADD45B with short interfering RNAs (siRNAs), as with miR-K9. KSHV miR-K9 also decreased the protein levels of cleaved caspase-3, cleaved caspase-7, and cleaved poly(ADP-ribose) polymerase (PARP). In B lymphocytes latently infected with KSHV, specific inhibitors of KSHV miR-K9 led to increased GADD45B expression and apoptosis, indicating that miR-K9 is important for reducing apoptosis in infected cells. Furthermore, ectopic expression of GADD45B in KSHV-infected cells promoted apoptosis. Together, these results identify a new miRNA target and demonstrate that KSHV miRNAs are important for protecting infected cells from DNA damage responses. IMPORTANCE Kaposi's sarcoma-associated herpesvirus is a leading cause of cancers in individuals with AIDS. Promoting survival of infected cells is essential for maintaining viral infections. A virus needs to combat various cellular defense mechanisms designed to eradicate the viral infection. One such response can include DNA damage response factors, which can promote an arrest in cell growth and trigger cell death. We used a new approach to search for human genes repressed by small nucleic acids (microRNAs) expressed by a gammaherpesvirus (KSHV), which identified a gene called GADD45B as a target of microRNAs. Repression of GADD45B, which is expressed in response to DNA damage, benefited survival of infected cells in response to a DNA damage response. This information could be used to design new treatments for herpesvirus infections.

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Section: Resultssupporting

confidence: 52%

Section: Resultsmentioning

confidence: 99%

Kaposi's Sarcoma-Associated Herpesvirus MicroRNAs Target GADD45B To Protect Infected Cells from Cell Cycle Arrest and Apoptosis

Liu

Happel

Ziegelbauer

2017

J Virol

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“…More recently, miRNAs were shown to be derived from 12 precursor miRNAs (pre-miRNAs) encoded by sequences in the latency locus of the KSHV genome 60 . These miRNAs can contribute to oncogenesis by modulating cell differentiation 60 , cytokine production, and immune receptor signaling 61 .…”

Section: Etiologic Agents and Pathogenesismentioning

confidence: 99%

Cancer prevention in HIV-infected populations

Gonçalves

Montezuma-Rusca

Yarchoan

et al. 2016

Seminars in Oncology

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People living with human immunodeficiency virus (HIV) are living longer since the advent of effective combined antiretroviral therapy (cART). While cART substantially decreases the risk of developing some cancers, HIV-infected individuals remain at high risk for Kaposi sarcoma, lymphoma and several solid tumors. Currently HIV-infected patients represent an aging group, and malignancies have become a leading cause of morbidity and mortality. Tailored cancer-prevention strategies are needed for this population. In this review we describe the etiologic agents and pathogenesis of common malignancies in the setting of HIV, as well as current evidence for cancer prevention strategies and screening programs.

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“…Furthermore, some miRNA targets are repressed at the level of translation and display little or no change at the mRNA level. Recently, a report have shown that using proteomic screening has been successful to identify viral miRNA targets that would be missed looking at the mRNA level [39][40]. Ideally, it is best to search for gene expression changes when showing a gain or loss of a single miRNA function.…”

Section: Mirna Target Prediction Strategiesmentioning

confidence: 99%

Viral microRNA genomics and target validation

Ziegelbauer¹

2014

Current Opinion in Virology

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A subset of viruses express their own microRNAs (miRNAs) and one way to understand the functions of these microRNAs is to identify the targets of these miRNAs. Sequence analysis and mRNA expression profiling were some of the first techniques to identify targets of viral miRNAs. More recently, proteomics and sequencing of RNA by crosslinking and immunoprecipitation (CLIP) methods have been insightful and discovered many miRNA targets that may be missed using other methods. We are now at a point where numerous validated miRNA targets have been described and integration of these genomic datasets will provide a richer understanding of miRNA targeting and viral infection, persistence, and pathogenesis.

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Proteomic Screening of Human Targets of Viral microRNAs Reveals Functions Associated with Immune Evasion and Angiogenesis

Cited by 52 publications

References 56 publications

Kaposi's Sarcoma-Associated Herpesvirus MicroRNAs Target GADD45B To Protect Infected Cells from Cell Cycle Arrest and Apoptosis

Kaposi's Sarcoma-Associated Herpesvirus MicroRNAs Target GADD45B To Protect Infected Cells from Cell Cycle Arrest and Apoptosis

Cancer prevention in HIV-infected populations

Viral microRNA genomics and target validation

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