Proteomics and Phosphoproteomics of an Inducible HIV

ABSTRACT: The mechanisms by which human immunodeficiency virus (HIV) circumvents cellular defense machinery to replicate and persist in cells are not fully understood. HIV accessory proteins play key roles in the HIV life cycle by altering host signaling pathways that are highly dependent on post-translational modification (PTM) events. Thus, the identification of HIV accessory protein host targets and their PTM status is critical to fully understand how HIV invades, avoids detection and replicates to spread infection. To date, a comprehensive characterization of HIV accessory protein host targets and modulation of their PTM status does not exist. The significant gap in knowledge regarding the identity and PTMs of HIV host targets is due, in part, to technological limitations. Through this study we sought to apply current mass spectrometry techniques to define mechanisms of viral protein action through identifying host protein interaction partners of Vpr and modulation of down-stream PTM based signaling pathways. Here we report identification and abundance dynamics of over 7,000 proteins and 28,000 phospho-peptides. By utilizing a novel, inducible HIV-1 CD4+ T-cell model system, we overcame challenges associated with synchronization and infection present in other models. This inducible HIV-1 model, in conjunction with state of the art quantitative multiplexed proteomics, allowed for accurate assessment of early protein and PTM dynamics associated with induction of HIV expression. This thorough characterization of Vpr interactions and PTM temporal dynamics, assessed by comparing the wild type with a Vpr deficient strain, deepens the understanding of how HIV circumvents immunity to hijack host cells. This study acts as a resource that lays the foundation for validating host proteins and important PTM based signaling pathways as viable intervention targets.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Hiv-1 Unknown Group (ncbitaxon:540993) Homo Sapiens (ncbitaxon:9606)

SUBMITTER: David Gonzalez  

PROVIDER: MSV000080877 | MassIVE | Sat Apr 01 16:07:00 BST 2017

SECONDARY ACCESSION(S): PXD006230

REPOSITORIES: MassIVE