PMID- 25496667
OWN - NLM
STAT- MEDLINE
DCOM- 20151012
LR  - 20220318
IS  - 1742-4690 (Electronic)
IS  - 1742-4690 (Linking)
VI  - 11
DP  - 2014 Dec 13
TI  - Genome-wide shRNA screening identifies host factors involved in early endocytic 
      events for HIV-1-induced CD4 down-regulation.
PG  - 118
LID - 10.1186/s12977-014-0118-4 [doi]
LID - 118
AB  - BACKGROUND: Down-modulation of the CD4 receptor is one of the hallmarks of HIV-1 
      infection and it is believed to confer a selective replicative advantage to the 
      virus in vivo. This process is mainly mediated by three viral proteins: Env, Vpu 
      and Nef. To date, the mechanisms that lead to CD4 depletion from the surface of 
      infected cells during HIV-1 infection are still only partially characterized. In 
      this study, we sought to identify and characterize cellular host factors in 
      HIV-1-induced CD4 down-modulation. RESULTS: To identify host factors involved in 
      CD4 down-regulation, we used a whole genome-targeting shRNA lentiviral library in 
      HeLa CD4+ cells expressing Nef as an inducer of CD4 down-modulation. We 
      identified 55 genes, mainly encoding for proteins involved in various steps of 
      clathrin-mediated endocytosis. For confirmation and further selection of the hits 
      we performed several rounds of validation, using individual shRNA lentiviral 
      vectors with a different target sequence for gene knock-down in HIV-1-infected T 
      cells. By this stringent validation set-up, we could demonstrate that the 
      knock-down of DNM3 (dynamin 3), SNX22 (sorting nexin 22), ATP6AP1 (ATPase, H+ 
      Transporting, Lysosomal Accessory Protein 1), HRBL (HIV-Rev binding protein 
      Like), IDH3G (Isocitrate dehydrogenase), HSP90B1 (Heat shock protein 90 kDa beta 
      member 1) and EPS15 (Epidermal Growth Factor Receptor Pathway Substrate 15) 
      significantly increases CD4 levels in HIV-infected SupT1 T cells compared to the 
      non-targeting shRNA control. Moreover, EPS15, DNM3, IDH3G and ATP6AP1 knock-down 
      significantly decreases HIV-1 replication in T cells. CONCLUSIONS: We identified 
      seven genes as cellular co-factors for HIV-1-mediated CD4 down-regulation in T 
      cells. The knock-down of four out of seven of these genes also significantly 
      reduces HIV-1 replication in T cells. Next to a role in HIV-mediated CD4 
      down-regulation, these genes might however affect HIV-1 replication in another 
      way. Our findings give insights in the HIV-1-mediated CD4 down-regulation at the 
      level of the plasma membrane and early endosomes and identify four possible new 
      HIV-1 replication co-factors.
FAU - Landi, Alessia
AU  - Landi A
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      alessia.landi@ugent.be.
FAU - Vermeire, Jolien
AU  - Vermeire J
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      jolien.vermeire@ugent.be.
FAU - Iannucci, Veronica
AU  - Iannucci V
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      veronica.iannucci@mssm.edu.
AD  - Present address: Department of Microbiology, Icahn School of Medicine at Mount 
      Sinai, New York, NY, USA. veronica.iannucci@mssm.edu.
FAU - Vanderstraeten, Hanne
AU  - Vanderstraeten H
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      hanne.vanderstraeten@ugent.be.
FAU - Naessens, Evelien
AU  - Naessens E
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      evelien.naessens@ugent.be.
FAU - Bentahir, Mostafa
AU  - Bentahir M
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      mostafa.bentahir@uclouvain.be.
AD  - Present address: Centre de Technologies Moleculaires Appliquees, Ecole de Sante 
      Publique, Brussels, Belgium. mostafa.bentahir@uclouvain.be.
FAU - Verhasselt, Bruno
AU  - Verhasselt B
AD  - Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University 
      and Ghent University Hospital, De Pintelaan 185, B-9000, Gent, Belgium. 
      bruno.verhasselt@ugent.be.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20141213
PL  - England
TA  - Retrovirology
JT  - Retrovirology
JID - 101216893
RN  - 0 (CD4 Antigens)
RN  - 0 (RNA, Small Interfering)
SB  - IM
MH  - CD4 Antigens/*biosynthesis
MH  - CD4-Positive T-Lymphocytes/*immunology/*virology
MH  - Cell Line
MH  - Down-Regulation
MH  - *Endocytosis
MH  - *Gene Expression Regulation
MH  - Genetic Testing
MH  - HIV-1/*immunology/*physiology
MH  - Host-Pathogen Interactions
MH  - Humans
MH  - RNA, Small Interfering/genetics
MH  - Virus Replication
PMC - PMC4269872
EDAT- 2014/12/17 06:00
MHDA- 2015/10/13 06:00
PMCR- 2014/12/13
CRDT- 2014/12/16 06:00
PHST- 2014/08/14 00:00 [received]
PHST- 2014/12/01 00:00 [accepted]
PHST- 2014/12/16 06:00 [entrez]
PHST- 2014/12/17 06:00 [pubmed]
PHST- 2015/10/13 06:00 [medline]
PHST- 2014/12/13 00:00 [pmc-release]
AID - s12977-014-0118-4 [pii]
AID - 118 [pii]
AID - 10.1186/s12977-014-0118-4 [doi]
PST - epublish
SO  - Retrovirology. 2014 Dec 13;11:118. doi: 10.1186/s12977-014-0118-4.