A genome-wide CRISPR screen identifies UFMylation and TRAMP-like complexes as host factors required for hepatitis A virus infection

Kulsuptrakul, Jessie; Wang, Ruofan; Meyers, Nathan L.; Ott, Melanie; Puschnik, Andreas S.

doi:10.1016/j.celrep.2021.108859

Cited by 37 publications

(27 citation statements)

References 78 publications

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“…To confirm the existence of additional peptidases with activity towards precursor UFM1, we generated UFSP2 −/− cell lines using CRISPR-Cas9 approaches. Consistent with previous studies, we observed an increase in both RPL26-(UFM1)1 and RPL26-(UFM1)2 species as a result of UFSP2 deficiency ( Fig-1A , Fig-S1A-D ) (Walczak et al , 2019; Liang et al , 2020; Wang et al , 2020; Kulsuptrakul et al , 2021). We next developed an experimental system to monitor UFM1 peptidase activity, where cell lysates were incubated with a reporter protein comprising pro-UFM1 1-85 fused at its carboxy-terminus to GFP via a short peptide linker.…”

Section: Resultssupporting

confidence: 92%

Human UFSP1 is an active protease that regulates UFM1 maturation and UFMylation

Millrine

Cummings

Matthews

et al. 2022

Preprint

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An essential first step in the posttranslational modification of proteins with UFM1, UFMylation, is the proteolytic cleavage of pro-UFM1 to expose a C-terminal glycine. Of the two UFM1-specific proteases (UFSPs) identified in humans, only UFSP2 is reported to be active since the annotated sequence of UFSP1 lacks critical catalytic residues. Nonetheless, efficient UFM1 maturation occurs in cells lacking UFSP2 suggesting the presence of another active protease. We hereby identify a long isoform of UFSP1 to be this protease. Cells lacking both UFSPs show complete loss of UFMylation resulting from an absence of mature UFM1. While UFSP2, but not UFSP1, removes UFM1 from the ribosomal subunit RPL26, UFSP1 acts earlier in the pathway to mature UFM1 and cleave a potential auto-inhibitory modification on UFC1, thereby controlling activation of UFMylation. In summary, our studies reveal important distinctions in substrate specificity and localization-dependent functions for the two proteases in regulating UFMylation.

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Section: Resultssupporting

confidence: 92%

Human UFSP1 is an active protease that regulates UFM1 maturation and UFMylation

Millrine

Cummings

Matthews

et al. 2022

Preprint

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“…The methods used can be multiple and complementary, spanning from microscopy techniques [41] to in silico modeling [4,42,43]; however, here, we focus on the enzymatic, pharmacological, and structural approaches.…”

Section: Confirmatory Methods: Verification Of the Glycan-virus Interactionmentioning

confidence: 99%

Viruses Like Sugars: How to Assess Glycan Involvement in Viral Attachment

Mathez

Cagno

2021

Microorganisms

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The first step of viral infection requires interaction with the host cell. Before finding the specific receptor that triggers entry, the majority of viruses interact with the glycocalyx. Identifying the carbohydrates that are specifically recognized by different viruses is important both for assessing the cellular tropism and for identifying new antiviral targets. Advances in the tools available for studying glycan–protein interactions have made it possible to identify them more rapidly; however, it is important to recognize the limitations of these methods in order to draw relevant conclusions. Here, we review different techniques: genetic screening, glycan arrays, enzymatic and pharmacological approaches, and surface plasmon resonance. We then detail the glycan interactions of enterovirus D68 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), highlighting the aspects that need further clarification.

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“…SARs-CoV-2 Nsp1•40S interactions may also prove amenable to antiviral drug development. Cellular decay factors, including the SKI and TRAMP-like complex, and m 6 A modification by METTL3 on which viruses depend but whose short-term inhibition is tolerated by the host also present druggable targets less likely to be overcome by virus resistance ( Weston et al 2020 ; Burgess et al 2021 ; Ho et al 2021 ; Kulsuptrakul et al 2021 ). By tinkering with functions that regulate eIF2α phosphorylation, a safe, attenuated HSV-1 that preferentially replicates in cancer cells with impaired cell-intrinsic immune responses was engineered ( Taneja et al 2001 ).…”

Section: Closing Thoughtsmentioning

confidence: 99%

“…Finally, ubiquitin fold modifier (UFM1) conjugation to uL24 (RPL26) ( Fig. 4 ) is required for HAV translation ( Kulsuptrakul et al 2021 ).…”

Section: Appropriating Host Ribosomes In Virus-infected Cellsmentioning

confidence: 99%

Minding the message: tactics controlling RNA decay, modification, and translation in virus-infected cells

2022

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With their categorical requirement for host ribosomes to translate mRNA, viruses provide a wealth of genetically tractable models to investigate how gene expression is remodeled post-transcriptionally by infection-triggered biological stress. By co-opting and subverting cellular pathways that control mRNA decay, modification, and translation, the global landscape of post-transcriptional processes is swiftly reshaped by virus-encoded factors. Concurrent host cell-intrinsic countermeasures likewise conscript post-transcriptional strategies to mobilize critical innate immune defenses. Here we review strategies and mechanisms that control mRNA decay, modification, and translation in animal virus-infected cells. Besides settling infection outcomes, post-transcriptional gene regulation in virus-infected cells epitomizes fundamental physiological stress responses in health and disease.

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A genome-wide CRISPR screen identifies UFMylation and TRAMP-like complexes as host factors required for hepatitis A virus infection

Cited by 37 publications

References 78 publications

Human UFSP1 is an active protease that regulates UFM1 maturation and UFMylation

Human UFSP1 is an active protease that regulates UFM1 maturation and UFMylation

Viruses Like Sugars: How to Assess Glycan Involvement in Viral Attachment

Minding the message: tactics controlling RNA decay, modification, and translation in virus-infected cells

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