CRISPR-based gene knockout screens reveal deubiquitinases involved in HIV-1 latency in two Jurkat cell models.
CRISPR-Cas Systems
DNA Polymerase III
/ genetics
DNA-Binding Proteins
/ genetics
DNA-Directed RNA Polymerases
/ genetics
Deubiquitinating Enzyme CYLD
/ genetics
Deubiquitinating Enzymes
/ antagonists & inhibitors
Endopeptidases
/ genetics
Enzyme Inhibitors
/ pharmacology
Gene Knockout Techniques
/ methods
HIV-1
/ genetics
Host-Pathogen Interactions
/ genetics
Humans
Jurkat Cells
Nucleoproteins
/ genetics
Proteasome Endopeptidase Complex
/ genetics
RNA-Binding Proteins
/ genetics
Transcription Factors
/ genetics
Ubiquitin Thiolesterase
/ genetics
Virus Latency
/ drug effects
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
24 03 2020
24 03 2020
Historique:
received:
31
10
2019
accepted:
10
03
2020
entrez:
27
3
2020
pubmed:
27
3
2020
medline:
19
12
2020
Statut:
epublish
Résumé
The major barrier to a HIV-1 cure is the persistence of latent genomes despite treatment with antiretrovirals. To investigate host factors which promote HIV-1 latency, we conducted a genome-wide functional knockout screen using CRISPR-Cas9 in a HIV-1 latency cell line model. This screen identified IWS1, POLE3, POLR1B, PSMD1, and TGM2 as potential regulators of HIV-1 latency, of which PSMD1 and TMG2 could be confirmed pharmacologically. Further investigation of PSMD1 revealed that an interacting enzyme, the deubiquitinase UCH37, was also involved in HIV-1 latency. We therefore conducted a comprehensive evaluation of the deubiquitinase family by gene knockout, identifying several deubiquitinases, UCH37, USP14, OTULIN, and USP5 as possible HIV-1 latency regulators. A specific inhibitor of USP14, IU1, reversed HIV-1 latency and displayed synergistic effects with other latency reversal agents. IU1 caused degradation of TDP-43, a negative regulator of HIV-1 transcription. Collectively, this study is the first comprehensive evaluation of deubiquitinases in HIV-1 latency and establishes that they may hold a critical role.
Identifiants
pubmed: 32210344
doi: 10.1038/s41598-020-62375-3
pii: 10.1038/s41598-020-62375-3
pmc: PMC7093534
doi:
Substances chimiques
DNA-Binding Proteins
0
Enzyme Inhibitors
0
Iws1 protein, human
0
Nucleoproteins
0
POLE3 protein, human
0
PSMD1 protein, human
0
RNA-Binding Proteins
0
TARDBP protein, human
0
Transcription Factors
0
USP14 protein, human
0
DNA-Directed RNA Polymerases
EC 2.7.7.6
DNA Polymerase III
EC 2.7.7.7
Endopeptidases
EC 3.4.-
CYLD protein, human
EC 3.4.19.12
Deubiquitinating Enzyme CYLD
EC 3.4.19.12
Deubiquitinating Enzymes
EC 3.4.19.12
UCHL5 protein, human
EC 3.4.19.12
Ubiquitin Thiolesterase
EC 3.4.19.12
Proteasome Endopeptidase Complex
EC 3.4.25.1
ubiquitin isopeptidase
EC 3.4.99.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
5350Subventions
Organisme : NIAID NIH HHS
ID : R01 AI134328
Pays : United States
Organisme : NIAID NIH HHS
ID : T32 AI106711
Pays : United States
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