Novel HIV-1 knockdown targets identified by an enriched kinases/phosphatases shRNA library using a long-term iterative screen in Jurkat T-cells.
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.
Rato
Sylvie
S
URIA-Centro de Patogénese Molecular, Faculdade de Farmácia, Universidade de Lisboa, Lisboa, Portugal.
Maia
Sara
S
Brito
Paula M
PM
Resende
Leonor
L
Pereira
Carina F
CF
Moita
Catarina
C
Freitas
Rui P
RP
Moniz-Pereira
José
J
Hacohen
Nir
N
Moita
Luis Ferreira
LF
Goncalves
Joao
J
D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival; D015723:Gene Library; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D019169:Jurkat Cells; D015458:Leukemia, T-Cell / Q000235:genetics / Q000473:pathology / Q000821:virology; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics* / Q000378:metabolism; D010770:Phosphotransferases / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology; D054320:vif Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism
D034741:RNA, Small Interfering; D054320:vif Gene Products, Human Immunodeficiency Virus; C514847:vif protein, Human immunodeficiency virus 1; D010770:Phosphotransferases; D010744:Phosphoric Monoester Hydrolases