title: A functional role for ADAM10 in human immunodeficiency virus type-1 replication.
abstract: BACKGROUND
  Gene trap insertional mutagenesis was used as a high-throughput approach to discover cellular genes participating in viral infection by screening libraries of cells selected for survival from lytic infection with a variety of viruses. Cells harboring a disrupted ADAM10 (A Disintegrin and Metalloprotease 10) allele survived reovirus infection, and subsequently ADAM10 was shown by RNA interference to be important for replication of HIV-1.
  
  
  RESULTS
  Silencing ADAM10 expression with small interfering RNA (siRNA) 48 hours before infection significantly inhibited HIV-1 replication in primary human monocyte-derived macrophages and in CD4⁺ cell lines. In agreement, ADAM10 over-expression significantly increased HIV-1 replication. ADAM10 down-regulation did not inhibit viral reverse transcription, indicating that viral entry and uncoating are also independent of ADAM10 expression. Integration of HIV-1 cDNA was reduced in ADAM10 down-regulated cells; however, concomitant 2-LTR circle formation was not detected, suggesting that HIV-1 does not enter the nucleus. Further, ADAM10 silencing inhibited downstream reporter gene expression and viral protein translation. Interestingly, we found that while the metalloprotease domain of ADAM10 is not required for HIV-1 replication, ADAM15 and γ-secretase (which proteolytically release the extracellular and intracellular domains of ADAM10 from the plasma membrane, respectively) do support productive infection.
  
  
  CONCLUSIONS
  We propose that ADAM10 facilitates replication at the level of nuclear trafficking. Collectively, our data support a model whereby ADAM10 is cleaved by ADAM15 and γ-secretase and that the ADAM10 intracellular domain directly facilitates HIV-1 nuclear trafficking. Thus, ADAM10 represents a novel cellular target class for development of antiretroviral drugs.
authors.affiliation: Department of Pathology, University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA.
authors.forename: Brian M
authors.initials: BM
authors.lastname: Friedrich
authors.forename: James L
authors.initials: JL
authors.lastname: Murray
authors.forename: Guangyu
authors.initials: G
authors.lastname: Li
authors.forename: Jinsong
authors.initials: J
authors.lastname: Sheng
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Hodge
authors.forename: Donald H
authors.initials: DH
authors.lastname: Rubin
authors.forename: William A
authors.initials: WA
authors.lastname: O'Brien
authors.forename: Monique R
authors.initials: MR
authors.lastname: Ferguson
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072197:ADAM10 Protein; D021581:Active Transport, Cell Nucleus; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D002478:Cells, Cultured; D015497:HIV-1 / Q000472:pathogenicity* / Q000502:physiology; D054884:Host-Pathogen Interactions; D006801:Humans; D008264:Macrophages / Q000821:virology; D008565:Membrane Proteins / Q000378:metabolism*; D008954:Models, Biological; D016254:Mutagenesis, Insertional; D016662:Virus Integration; D014779:Virus Replication
chemical_list: D008565:Membrane Proteins; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human