Proteomic and biochemical analysis of purified human immunodeficiency virus type 1 produced from infected monocyte-derived macrophages.
Human immunodeficiency virus type 1 (HIV-1) infects CD4(+) T lymphocytes and monocytes/macrophages, incorporating host proteins in the process of assembly and budding. Analysis of the host cell proteins incorporated into virions can provide insights into viral biology. We characterized proteins in highly purified HIV-1 virions produced from human monocyte-derived macrophages (MDM), within which virus buds predominantly into intracytoplasmic vesicles, in contrast to the plasmalemmal budding of HIV-1 typically seen with infected T cells. Liquid chromatography-linked tandem mass spectrometry of highly purified virions identified many cellular proteins, including 33 previously described proteins in HIV-1 preparations from other cell types. Proteins involved in many different cellular structures and functions were present, including those from the cytoskeleton, adhesion, signaling, intracellular trafficking, chaperone, metabolic, ubiquitin/proteasomal, and immune response systems. We also identified annexins, annexin-binding proteins, Rab proteins, and other proteins involved in membrane organization, vesicular trafficking, and late endosomal function, as well as apolipoprotein E, which participates in cholesterol transport, immunoregulation, and modulation of cell growth and differentiation. Several tetraspanins, markers of the late endosomal compartment, were also identified. MDM-derived HIV contained 26 of 37 proteins previously found in exosomes, consistent with the idea that HIV uses the late endosome/multivesicular body pathway during virion budding from macrophages.
Chertova
Elena
E
AIDS Vaccine Research Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, MD 21702, USA. chertova@ncifcrf.gov
Chertov
Oleg
O
Coren
Lori V
LV
Roser
James D
JD
Trubey
Charles M
CM
Bess
Julian W
JW
Sowder
Raymond C
RC
Barsov
Eugene
E
Hood
Brian L
BL
Fisher
Robert J
RJ
Nagashima
Kunio
K
Conrads
Thomas P
TP
Veenstra
Timothy D
TD
Lifson
Jeffrey D
JD
Ott
David E
DE
D002460:Cell Line; D015699:HIV Envelope Protein gp120 / Q000737:chemistry; D015497:HIV-1 / Q000302:isolation & purification* / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D017493:Leukocyte Common Antigens / Q000096:biosynthesis; D008264:Macrophages / Q000166:cytology* / Q000821:virology; D009000:Monocytes / Q000166:cytology* / Q000821:virology; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000821:virology*; D014357:Trypsin / Q000494:pharmacology; D014764:Viral Proteins / Q000737:chemistry; D014779:Virus Replication
D015699:HIV Envelope Protein gp120; D014764:Viral Proteins; D017493:Leukocyte Common Antigens; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D014357:Trypsin