title: Identification of cellular proteins required for replication of human immunodeficiency virus type 1.
abstract: Cellular proteins are essential for human immunodeficiency virus type 1 (HIV-1) replication and may serve as viable new targets for treating infection. Using gene trap insertional mutagenesis, a high-throughput approach based on random inactivation of cellular genes, candidate genes were found that limit virus replication when mutated. Disrupted genes (N=87) conferring resistance to lytic infection with several viruses were queried for an affect on HIV-1 replication by utilizing small interfering RNA (siRNA) screens in TZM-bl cells. Several genes regulating diverse pathways were found to be required for HIV-1 replication, including DHX8, DNAJA1, GTF2E1, GTF2E2, HAP1, KALRN, UBA3, UBE2E3, and VMP1. Candidate genes were independently tested in primary human macrophages, toxicity assays, and/or Tat-dependent β-galactosidase reporter assays. Bioinformatics analyses indicated that several host factors present in this study participate in canonical pathways and functional processes implicated in prior genome-wide studies. However, the genes presented in this study did not share identity with those found previously. Novel antiviral targets identified in this study should open new avenues for mechanistic investigation.
authors.affiliation: Department of Internal Medicine, University of Texas Medical Branch, Galveston, USA.
authors.forename: Natallia
authors.initials: N
authors.lastname: Dziuba
authors.forename: Monique R
authors.initials: MR
authors.lastname: Ferguson
authors.forename: William A
authors.initials: WA
authors.lastname: O'Brien
authors.forename: Anthony
authors.initials: A
authors.lastname: Sanchez
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Prussia
authors.forename: Natalie J
authors.initials: NJ
authors.lastname: McDonald
authors.forename: Brian M
authors.initials: BM
authors.lastname: Friedrich
authors.forename: Guangyu
authors.initials: G
authors.lastname: Li
authors.forename: Michael W
authors.initials: MW
authors.lastname: Shaw
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: James L
authors.initials: JL
authors.lastname: Murray
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics; D015497:HIV-1 / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D008297:Male; D008403:Mass Screening; D016254:Mutagenesis, Insertional / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D014775:Virus Activation / Q000235:genetics* / Q000502:physiology; D014779:Virus Replication / Q000235:genetics* / Q000502:physiology
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral