Novel HIV-1 Knockdown Targets Identified by an Enriched Kinases/Phosphatases shRNA Library Using a Long-Term Iterative Screen in Jurkat T-Cells

Autor: Carina F. Pereira, Nir Hacohen, Luis F. Moita, Paula M. Brito, José Moniz-Pereira, Catarina Moita, Sylvie Rato, João Gonçalves, Leonor Resende, Rui P. Freitas, Sara Maia
Rok vydání: 2010
Předmět:
MAPK/ERK pathway
Leukemia
T-Cell
Cell Survival
DNA repair
Blotting
Western
lcsh:Medicine
Virus Replication
Jurkat cells
Cell Line
Small hairpin RNA
Jurkat Cells
03 medical and health sciences
Retrovirus
Infectious Diseases/Viral Infections
vif Gene Products
Human Immunodeficiency Virus
Humans
RNA
Small Interfering
Virology/Effects of Virus Infection on Host Gene Expression
lcsh:Science
Gene Library
030304 developmental biology
0303 health sciences
Multidisciplinary
biology
030306 microbiology
Kinase
Phosphotransferases
lcsh:R
Infectious Diseases/HIV Infection and AIDS
biology.organism_classification
Molecular biology
Phosphoric Monoester Hydrolases
3. Good health
Cell biology
Virology/Viral Replication and Gene Regulation
Viral replication
Virology/Immunodeficiency Viruses
Host-Pathogen Interactions
HIV-1
RNA Interference
lcsh:Q
Virology/Host Antiviral Responses
HeLa Cells
Research Article
Genetic screen
Zdroj: PLoS ONE, Vol 5, Iss 2, p e9276 (2010)
PLoS ONE
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0009276
Popis: HIV-1 is a complex retrovirus that uses host machinery to promote its replication. Understanding cellular proteins involved in the multistep process of HIV-1 infection may result in the discovery of more adapted and effective therapeutic targets. Kinases and phosphatases are a druggable class of proteins critically involved in regulation of signal pathways of eukaryotic cells. Here, we focused on the discovery of kinases and phosphatases that are essential for HIV-1 replication but dispensable for cell viability. We performed an iterative screen in Jurkat T-cells with a short-hairpin-RNA (shRNA) library highly enriched for human kinases and phosphatases. We identified 14 new proteins essential for HIV-1 replication that do not affect cell viability. These proteins are described to be involved in MAPK, JNK and ERK pathways, vesicular traffic and DNA repair. Moreover, we show that the proteins under study are important in an early step of HIV-1 infection before viral integration, whereas some of them affect viral transcription/translation. This study brings new insights for the complex interplay of HIV-1/host cell and opens new possibilities for antiviral strategies.
Databáze: OpenAIRE
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