Proteomic and Biochemical Analysis of Purified Human Immunodeficiency Virus Type 1 Produced from Infected Monocyte-Derived Macrophages

Abstract

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.As an RNA virus with limited coding capacity, human immunodeficiency virus type 1 (HIV-1) subverts cellular pathways and processes to facilitate many aspects of its replication cycle. It is known that a variety of cellular factors are involved in HIV-1 assembly and budding (13,20,42,44). Typically, HIV-1 is observed by electron microscopy to assemble at and bud from the plasma membrane in T cells and the epithelial cell lines that serve as models for HIV-1 assembly studies (23). In contrast, in macrophages, one of the primary target cell populations in vivo, HIV-1 appears to assemble mostly at internal late endosomal and multivesicular body (MVB) membranes and then bud into these vesicular structures, observable in electron micrographs as internal virion-filled compartments (48,54,55,59). After budding into MVB, these virion-laden vesicles are presumably transported to the cell surface and virus is released from the cell by a normal exocytotic fusion of these structures with the plasma membrane, thereby releasing the contents of the MVB (54, 55).To date, the differences in viral and cellular protein interactions involved in assembly and budding at the plasma membrane versus the late endosomal assembly pathway remain unclear. Clues to the location and the mechanism by which HIV-1 buds can be provided by the cellular proteins that are incorporated into virions. In the case of macrophage-derived virus, the presence of HLA class II and other late endosoma...

The Feline Calicivirus Leader of the Capsid Protein Is Associated with Cytopathic Effect

“…ANXA2 is a multifunctional protein and is involved in many cellular processes, such as fibrinolysis, exocytosis, endocytosis, cell-cell adhesion, and cell motility (36,37,48). ANXA2 has been linked to the life cycle of other positive-sense single-stranded RNA viruses, such as hepatitis C virus (49)(50)(51) and HIV (52)(53)(54). In both cases, a viral protein has been reported to interact with ANXA2 in infected cells, and silencing of ANXA2 affected the infectivity of the viruses.…”

Exosomes as intercellular signalosomes and pharmacological effectors

“…This could explain how viruses like HIV can cross the blood-brain barrier. HIV and exosome feature a similar size and share some protein content [82]; and HIV virions are partly Page 13 of 32 A c c e p t e d M a n u s c r i p t 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 13 generated into the MVB [82]. In addition recent developments demonstrated that HIV enters the endocytic track of target cells for infection [83].…”

“…When infected, dendritic cells, monocytes, macrophages and lymphocytes can produce both exosomes and HIV virions [82] with similar density, which require separation by immunocapture or appropriate gradient density separation. Also exosomes compete with HIV for the virus entry in endocytic compartment [83].…”

“…Also exosomes compete with HIV for the virus entry in endocytic compartment [83]. About 10% proteins are identical between HIV virions and exosomes issued from the same cell type [82].…”

Emerging functions as host cell factors – an encyclopedia of annexin-pathogen interactions

“…Annexin A1 (AnxA1) is one of the most famous family members and there is a wealth of information on its protective anti-inflammatory and proresolving effects (Perretti and D'Acquisto, 2009 (Kattenhorn et al, 2004;Dry et al, 2008;Loret et al, 2008;Vidick et al, 2013), human immunodeficiency virus type 1 (HIV-1) (Chertova et al, 2006), vesicular stomatitis virus (VSV) (Moerdyk-Schauwecker et al, 2009, Rift Valley fever virus (Nuss et al, 2014) and IAV (Shaw et al, 2008), and recent evidence points to a function of the AnxA1 receptor FPR2 in IAV replication (Tcherniuk et al, 2016). AnxA2 is a component of herpesviruses (Wright et al, 1994(Wright et al, , 1995Varnum et al, 2004;Zhu et al, 2005;Dry et al, 2008;Loret et al, 2008;Gershom et al, 2012;Vidick et al, 2013), IAV (LeBouder et al, 2008;Shaw et al, 2008;Liu et al, 2012), HIV-1 (Chertova et al, 2006), human papilloma virus (HPV) type 16 (Woodham et al, 2012), hepatitis C virus (HCV) type 1 (Backes et al, 2010), VSV (Moerdyk-Schauwecker et al, 2009, vaccinia virus (Chung et al, 2006), Rift Valley fever virus (Nuss et al, 2014) and Newcastle disease virus (Ren et al, 2012). AnxA5 is associated with herpes simplex virus (HSV) type 1 (Loret et al, 2008), HIV-1 (Chertova et al, 2006), VSV (Moerdyk-Schauwecker et al, 2009, Rift Valley fever virus (Nuss et al, 2014), and IAV (Shaw et al, 2008).…”