Replication of Merkel cell polyomavirus induces reorganization of promyelocytic leukemia nuclear bodies

Neumann, Friederike; Czech-Sioli, Manja; Dobner, Thomas; Grundhoff, Adam; Schreiner, Sabrina; Fischer, Nicole

doi:10.1099/jgv.0.000593

Cited by 13 publications

(12 citation statements)

References 52 publications

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“…Kap1 depletion increases MCPyV DNA replication. To evaluate the consequences of Kap1/T protein interaction, we performed MCPyV DNA replication assays using a semipermissive system (12,15,16,35,36) in cells positive or negative for Kap1 expression. We generated HEK293 cells ( Fig.…”

Section: Resultsmentioning

confidence: 99%

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Merkel Cell Polyomavirus DNA Replication Induces Senescence in Human Dermal Fibroblasts in a Kap1/Trim28-Dependent Manner

Siebels

Czech-Sioli

Spohn

et al. 2020

mBio

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Merkel cell polyomavirus (MCPyV) is the only polyomavirus known to be associated with tumorigenesis in humans. Similarly to other polyomaviruses, MCPyV expresses a large tumor antigen (LT-Ag) that, together with a small tumor antigen (sT-Ag), contributes to cellular transformation and that is of critical importance for the initiation of the viral DNA replication. Understanding the cellular protein network regulated by MCPyV early proteins will significantly contribute to our understanding of the natural MCPyV life cycle as well as of the mechanisms by which the virus contributes to cellular transformation. We here describe KRAB-associated protein 1 (Kap1), a chromatin remodeling factor involved in cotranscriptional regulation, as a novel protein interaction partner of MCPyV T antigens sT and LT. Kap1 knockout results in a significant increase in the level of viral DNA replication that is highly suggestive of Kap1 being an important host restriction factor during MCPyV infection. Differently from other DNA viruses, MCPyV gene expression is unaffected in the absence of Kap1 and Kap1 does not associate with the viral genome. Instead, we show that in primary normal human dermal fibroblast (nHDF) cells, MCPyV DNA replication, but not T antigen expression alone, induces ataxia telangiectasia mutated (ATM) kinase-dependent Kap1 S824 phosphorylation, a mechanism that typically facilitates repair of double-strand breaks in heterochromatin by arresting the cells in G2. We show that MCPyV-induced inhibition of cell proliferation is mainly conferred by residues within the origin binding domain and thereby by viral DNA replication. Our data suggest that phosphorylation of Kap1 and subsequent Kap1-dependent G2 arrest/senescence represent host defense mechanisms against MCPyV replication in nHDF cells. IMPORTANCE We here describe Kap1 as a restriction factor in MCPyV infection. We report a novel, indirect mechanism by which Kap1 affects MCPyV replication. In contrast with from other DNA viruses, Kap1 does not associate with the viral genome in MCPyV infection and has no impact on viral gene expression. In MCPyV-infected nHDF cells, Kap1 phosphorylation (pKap1 S824) accumulates because of genomic stress mainly induced by viral DNA replication. In contrast, ectopic expression of LT or LT MCPyV mutants, previously shown to be important for induction of genotoxic stress, does not result in a similar extent of pKap1 accumulation. We show that cells actively replicating MCPyV accumulate pKap1 (in a manner dependent on the presence of ATM) and display a senescence phenotype reflected by G2 arrest. These results are supported by transcriptome analyses showing that LT antigen, in a manner dependent on the presence of Kap1, induces expression of secreted factors, which is known as the senescence-associated secretory phenotype (SASP).

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Section: Resultsmentioning

confidence: 99%

“…MCPyV replication assay. In vitro replication assays were performed as previously described (15,16,35). For nHDF cells, 1 ϫ 10 6 cells were transfected with 2 g MCVSyn or pUC18 plasmid (mock) using electroporation.…”

Section: Methodsmentioning

confidence: 99%

Merkel Cell Polyomavirus DNA Replication Induces Senescence in Human Dermal Fibroblasts in a Kap1/Trim28-Dependent Manner

Siebels

Czech-Sioli

Spohn

et al. 2020

mBio

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“…While our RNA-seq data identified various cellular targets, many of which are involved in immune-related processes, it is important to acknowledge that given the HEK293-cell background, it is possible that some relevant targets were missed. SP100 is an intriguing MCV-miR-M1 target, which has been implicated in the innate immune response against double-stranded DNA viruses, including MCPyV (Gunther et al, 2014;Jiang et al, 2011;Neumann et al, 2016;Tavalai and Stamminger, 2009;Wagenknecht et al, 2015) and has been shown to be down-regulated during MCPyV replication, although no mechanism for this was reported (Neumann et al, 2016). Our data suggest that MCV-miR-M1 mediates, at least in part, the observed down-regulation of SP100 in cells harboring synthetic MCPyV.…”

Section: Discussionmentioning

confidence: 76%

MCV-miR-M1 Targets the Host-Cell Immune Response Resulting in the Attenuation of Neutrophil Chemotaxis

Akhbari

Tobin

Poterlowicz

et al. 2018

Journal of Investigative Dermatology

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Virus-encoded microRNAs are emerging as key regulators of persistent infection and host-cell immune evasion. Merkel cell polyomavirus, the predominant etiological agent of Merkel cell carcinoma, encodes a single microRNA, MCV-miR-M1, which targets the oncogenic Merkel cell polyomavirus large T antigen. MCV-miR-M1 has previously been shown to play an important role in the establishment of long-term infection, however, the underlying mechanism is not fully understood. A key unanswered question is whether, in addition to autoregulating large T antigen, MCV-miR-M1 also targets cellular transcripts to orchestrate an environment conducive to persistent infection. To address this, we adopted an RNA sequencing-based approach to identify cellular targets of MCV-miR-M1. Intriguingly, bioinformatics analysis of transcripts that are differentially expressed in cells expressing MCV-miR-M1 revealed several genes implicated in immune evasion. Subsequent target validation led to the identification of the innate immunity protein, SP100, as a direct target of MCV-miR-M1. Moreover, MCV-miR-M1-mediated modulation of SP100 was associated with a significant decrease in CXCL8 secretion, resulting in the attenuation of neutrophil chemotaxis toward Merkel cells harboring synthetic Merkel cell polyomavirus. Based on these observations, we propose that MCV-miR-M1 targets key immune response regulators to help facilitate persistent infection, which is a prerequisite for cellular transformation in Merkel cell carcinoma.

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“…In the maturation process they hydrolyse a peptide bond close to the C-terminus of SUMO precursors, eliminating the very C-terminal amino acids from SUMO1, SUMO2 and SUMO3 and exposing two glycine residues. In SUMO1-3, the diGly motif is preceded by a glutamine (Q) and threonine (T), while SUMO4 exhibits a PTGG motif, in which the proline residue confers resistance to SENPmediated cleavage (Owerbach et al, 2005). In the deconjugation process, SENPs cleave an isopeptide bond that links SUMO moieties to the ε-amino group of lysine residues.…”

Section: Senpsmentioning

confidence: 99%

“…MCPyV transforming ability mainly resides on the expression of Large T antigen proteins (Large-LT, and Short-ST) (Houben et al, 2010), even if molecular details driving MCPyV-mediated cell transformation are still not fully elucidated, although a role for ST is emerging (Shuda et al, 2011). Similarly, a role for viral exploitation of the SUMO pathway during Merkel cell carcinoma has not yet been investigated, even if a recent report shows that MCPyV replication depends on PML-NBs, suggesting a possible involvement of SUMOylation in regulating MCPyV transformation (Neumann et al, 2016).…”

Section: Epstein-barr Virusmentioning

confidence: 99%

Recent Highlights: Onco Viral Exploitation of the SUMO System

Mattoscio¹,

Medda²,

Chiocca³

2020

Current Issues in Molecular Biology

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Small ubiquitin-like modifier (SUMO)ylation is a crucial post-translational modification that controls functions of a wide collection of proteins and biological processes. Hence, given its pleiotropic role, viruses have developed many approaches to usurp SUMO conjugation to exploit the cellular host environment for their own benefit. Consistently, cancer cells also frequently impact on SUMO to force cellular transformation, underlining the importance of SUMO in health and diseases. Therefore, after a brief introduction to the multistep SUMOylation pathway, in this review we will focus our attention on several examples of strategies adopted by oncogenic viruses to hijack SUMOylation in order to promote infection, persistence and malignant transformation of host cells.caister.com/cimb Curr. Figure 3.1 The SUMO conjugation system. Ulp1/SENP proteases catalyse cleavage of the C-terminal domain of SUMO proteins, exposing a diglycine motif.Processed SUMO is transferred to a cysteine of the heterodimeric E1 enzyme Uba2/SAE1. SUMO is then conjugated to a cysteine of UBC9, the E2 enzyme and attached to a lysine residue of the consensus motif on target proteins. The conjugation is often facilitated by an E3-ligase, which enforces the interactions among the involved components. SUMOylation is a reversible pathway where the Ulp1/SENPs proteases dictate the de-SUMOylation process.caister.com/cimb 2 Curr. Issues Mol. Biol. Vol. 35Oncoviruses and SUMOylation | 37

show abstract

Replication of Merkel cell polyomavirus induces reorganization of promyelocytic leukemia nuclear bodies

Cited by 13 publications

References 52 publications

Merkel Cell Polyomavirus DNA Replication Induces Senescence in Human Dermal Fibroblasts in a Kap1/Trim28-Dependent Manner

Merkel Cell Polyomavirus DNA Replication Induces Senescence in Human Dermal Fibroblasts in a Kap1/Trim28-Dependent Manner

MCV-miR-M1 Targets the Host-Cell Immune Response Resulting in the Attenuation of Neutrophil Chemotaxis

Recent Highlights: Onco Viral Exploitation of the SUMO System

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