Productive herpesvirus lytic replication in primary effusion lymphoma cells requires S-phase entry

Hollingworth, Robert M.; Stewart, Grant S.; Grand, Roger J. A.

doi:10.1099/jgv.0.001444

Cited by 6 publications

(5 citation statements)

References 32 publications

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“…We demonstrate through flow cytometry analysis that the circHIPK3:miR‐30c:DLL4 circuit has a role in regulating the host cell cycle during KSHV replication. Although the role of KSHV in the cell cycle during latency is understood, with v‐cyclin playing a key role, the role of the cell cycle during KSHV lytic replication is debated, with lytic replication either requiring entry into S phase or G1 arrest (Wu et al , 2003; Jones et al , 2014; Hollingworth et al , 2020). Our results show that levels of cells in G1 phase slightly increase from 0 to 24 h while S phase is not significantly altered.…”

Section: Discussionmentioning

confidence: 99%

Dysregulation of the miR‐30c/DLL4 axis by circHIPK3 is essential for KSHV lytic replication

et al. 2022

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Non‐coding RNA (ncRNA) regulatory networks are emerging as critical regulators of gene expression. These intricate networks of ncRNA:ncRNA interactions modulate multiple cellular pathways and impact the development and progression of multiple diseases. Herpesviruses, including Kaposi’s sarcoma‐associated herpesvirus, are adept at utilising ncRNAs, encoding their own as well as dysregulating host ncRNAs to modulate virus gene expression and the host response to infection. Research has mainly focused on unidirectional ncRNA‐mediated regulation of target protein‐coding transcripts; however, we identify a novel host ncRNA regulatory network essential for KSHV lytic replication in B cells. KSHV‐mediated upregulation of the host cell circRNA, circHIPK3, is a key component of this network, functioning as a competing endogenous RNA of miR‐30c, leading to increased levels of the miR‐30c target, DLL4. Dysregulation of this network highlights a novel mechanism of cell cycle control during KSHV lytic replication in B cells. Importantly, disruption at any point within this novel ncRNA regulatory network has a detrimental effect on KSHV lytic replication, highlighting the essential nature of this network and potential for therapeutic intervention.

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Section: Discussionmentioning

confidence: 99%

Dysregulation of the miR‐30c/DLL4 axis by circHIPK3 is essential for KSHV lytic replication

et al. 2022

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“…Controls included uninfected SLK cells that were treated similarly which ruled out potential effects of chemical treatments (Supplementary Figure S1) as well as uninduced iSLK cells that were not treated with Dox and n-But. As in other experimental cell models of KSHV [75][76][77], the induction of lytic reactivation in infected iSLK cells is fractionally and asynchronous, and therefore, when examining cells post induction, we observed cells that did not undergo viral lytic reactivation as well as cells at different stages of the lytic cycle. These stages were distinguished by the intensity of the expression and the distribution of the lytic viral protein ORF45, which is predominantly nuclear at early lytic stages and perinuclear, cytoplasmic and close to the plasma membrane at later stages [62].…”

Section: Lytic Reactivation Of Kshv Is Associated With Major Changes ...mentioning

confidence: 52%

Lytic Reactivation of the Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) Is Accompanied by Major Nucleolar Alterations

Atari¹,

Rajan²,

Chikne³

et al. 2022

Viruses

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The nucleolus is a subnuclear compartment whose primary function is the biogenesis of ribosomal subunits. Certain viral infections affect the morphology and composition of the nucleolar compartment and influence ribosomal RNA (rRNA) transcription and maturation. However, no description of nucleolar morphology and function during infection with Kaposi’s sarcoma-associated herpesvirus (KSHV) is available to date. Using immunofluorescence microscopy, we documented extensive destruction of the nuclear and nucleolar architecture during the lytic reactivation of KSHV. This was manifested by the redistribution of key nucleolar proteins, including the rRNA transcription factor UBF. Distinct delocalization patterns were evident; certain nucleolar proteins remained together whereas others dissociated, implying that nucleolar proteins undergo nonrandom programmed dispersion. Significantly, the redistribution of UBF was dependent on viral DNA replication or late viral gene expression. No significant changes in pre-rRNA levels and no accumulation of pre-rRNA intermediates were found by RT-qPCR and Northern blot analysis. Furthermore, fluorescent in situ hybridization (FISH), combined with immunofluorescence, revealed an overlap between Fibrillarin and internal transcribed spacer 1 (ITS1), which represents the primary product of the pre-rRNA, suggesting that the processing of rRNA proceeds during lytic reactivation. Finally, small changes in the levels of pseudouridylation (Ψ) and 2′-O-methylation (Nm) were documented across the rRNA; however, none were localized to the functional domain. Taken together, our results suggest that despite dramatic changes in the nucleolar organization, rRNA transcription and processing persist during lytic reactivation of KSHV. Whether the observed nucleolar alterations favor productive infection or signify cellular anti-viral responses remains to be determined.

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“…We demonstrate through flow cytometry analysis the circHIPK3:miR-30c:DLL4 circuit has a role in regulating the host cell cycle during KSHV replication. Although the role of KSHV in the cell cycle during latency is understood, with v-Cyclin playing a key role, the role of the cell cycle during KSHV lytic replication is debated, with lytic replication either requiring entry into S phase or G1 arrest [49] [50] [51]. Our results show that levels of cells in G1 phase slightly increase from 0 to 24 hours while S phase is not significantly altered.…”

Section: Discussionmentioning

confidence: 82%

CircHIPK3 dysregulation of the miR-30c/DLL4 axis is essential for KSHV lytic replication

Harper

Mottram

Anene

et al. 2021

Preprint

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Non coding RNA (ncRNA) regulatory networks are emerging as critical regulators of gene expression. These intricate networks of ncRNA-ncRNA interactions modulate multiple cellular pathways and impact the development and progression of multiple diseases. Herpesviruses, including Kaposi's sarcoma-associated herpesvirus, are adept at utilising ncRNAs, encoding their own as well as dysregulating host ncRNAs to modulate virus gene expression and the host response to infection. Research has mainly focused on unidirectional ncRNA-mediated regulation of target protein-coding transcripts; however, we have identified a novel host ncRNA regulatory network essential for KSHV lytic replication in B cells. KSHV-mediated upregulation of the host cell circRNA, circHIPK3, is a key component of this network, functioning as a competing endogenous RNA of miR-30c, leading to increased levels of the miR-30c target, DLL4. Dysregulation of this network highlights a novel mechanism of cell cycle control during KSHV lytic replication in B cells. Importantly, disruption at any point within this novel ncRNA regulatory network has a detrimental effect on KSHV lytic replication, highlighting the essential nature of this network and potential for therapeutic intervention.

show abstract

Productive herpesvirus lytic replication in primary effusion lymphoma cells requires S-phase entry

Cited by 6 publications

References 32 publications

Dysregulation of the miR‐30c/DLL4 axis by circHIPK3 is essential for KSHV lytic replication

Dysregulation of the miR‐30c/DLL4 axis by circHIPK3 is essential for KSHV lytic replication

Lytic Reactivation of the Kaposi’s Sarcoma-Associated Herpesvirus (KSHV) Is Accompanied by Major Nucleolar Alterations

CircHIPK3 dysregulation of the miR-30c/DLL4 axis is essential for KSHV lytic replication

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