title: HIV-host interactome revealed directly from infected cells.
abstract: Although genetically compact, HIV-1 commandeers vast arrays of cellular machinery to sustain and protect it during cycles of viral outgrowth. Transposon-mediated saturation linker scanning mutagenesis was used to isolate fully replication-competent viruses harbouring a potent foreign epitope tag. Using these viral isolates, we performed differential isotopic labelling and affinity-capture mass spectrometric analyses on samples obtained from cultures of human lymphocytes to classify the vicinal interactomes of the viral Env and Vif proteins as they occur during natural infection. Importantly, interacting proteins were recovered without bias, regardless of their potential for positive, negative or neutral impact on viral replication. We identified specific host associations made with trimerized Env during its biosynthesis, at virological synapses, with innate immune effectors (such as HLA-E) and with certain cellular signalling pathways (for example, Notch1). We also defined Vif associations with host proteins involved in the control of nuclear transcription and nucleoside biosynthesis as well as those interacting stably or transiently with the cytoplasmic protein degradation apparatus. Our approach is broadly applicable to elucidating pathogen-host interactomes, providing high-certainty identification of interactors by their direct access during cycling infection. Understanding the pathophysiological consequences of these associations is likely to provide strategic targets for antiviral intervention.
authors.affiliation: Aaron Diamond AIDS Research Center, 455 1st Avenue, New York, New York 10016, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA.
authors.forename: Erica Y
authors.initials: EY
authors.lastname: Jacobs
authors.affiliation: Department of Molecular Biology, Lewis Thomas Laboratory, Princeton University, Princeton, New Jersey 08540, USA.
authors.forename: Todd M
authors.initials: TM
authors.lastname: Greco
authors.affiliation: Aaron Diamond AIDS Research Center, 455 1st Avenue, New York, New York 10016, USA.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Mohammed
authors.affiliation: San Diego Biomedical Research Institute, 10865 Road to the Cure, San Diego, California 92121, USA.
authors.forename: Tommy
authors.initials: T
authors.lastname: Tong
authors.affiliation: Department of Biochemistry, New York University Langone Medical Center, 227 East 30th Street, New York, New York 10016, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Keegan
authors.affiliation: San Diego Biomedical Research Institute, 10865 Road to the Cure, San Diego, California 92121, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Binley
authors.affiliation: Department of Molecular Biology, Lewis Thomas Laboratory, Princeton University, Princeton, New Jersey 08540, USA.
authors.forename: Ileana M
authors.initials: IM
authors.lastname: Cristea
authors.affiliation: Department of Biochemistry, New York University Langone Medical Center, 227 East 30th Street, New York, New York 10016, USA.
authors.forename: David
authors.initials: D
authors.lastname: Fenyö
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
authors.affiliation: Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA.
authors.forename: Brian T
authors.initials: BT
authors.lastname: Chait
authors.affiliation: Aaron Diamond AIDS Research Center, 455 1st Avenue, New York, New York 10016, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Muesing