PNNL

Abstract

Relatively little is known at the functional genomic level about the global host response to HIV-1 infection. Microarray analyses by several laboratories, including our own, have revealed that human immunodeficiency virus type 1 infection causes significant changes in host mRNA abundance and regulation of several cellular biological pathways. However, it remains unclear what consequences these changes bring about at the protein level. Here we report the expression levels of ~3,200 proteins assessed in the CD4+ CEMx174 cell line after infection with HIV-1 LAI, using liquid chromatography-mass spectrometry coupled with stable isotope labeling and the accurate mass and time (AMT) tag approach. Further, we found that 687 (21%) proteins changed in abundance at the peak of virus production at 36h post-infection. Pathway analysis revealed that the differential expression of proteins were concentrated in select biological pathways, exemplified by ubiquitin conjugating enzymes in the ubiquitination, carrier proteins in nucleo-cytoplasmic transport, cyclin-dependent kinase in cell cycle progression, and pyruvate dehydrogenase of the citrate cycle. Moreover, we observed changes in the abundance of proteins with known interactions with HIV-1 viral proteins. Our proteomic analysis captured changes in the host protein milieu at the time of robust virus production, accompanied by a moderate accumulation of G1/G0-phase cells. We will discuss the contributions of these changes to virus production in the infected cells.