Abstract
Cholesterol is an essential component of the cellular membranes and, by extension, of the HIV envelope membrane, which is derived from the host cell plasma membrane. Depletion of the cellular cholesterol has an inhibitory effect on HIV assembly, reduces infectivity of the produced virions, and makes the cell less susceptible to HIV infection. It is not surprising that the virus has evolved to gain access to cellular proteins regulating cholesterol metabolism. One of the key mechanisms used by HIV to maintain high levels of cholesterol in infected cells is Nef-mediated inhibition of cholesterol efflux and the cholesterol transporter responsible for this process, ABCA1. In this chapter, we describe methods to investigate these effects of HIV-1 infection.
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Waheed AA, Freed EO (2009) Lipids and membrane microdomains in HIV-1 replication. Virus Res 143:162–176
Ono A, Freed EO (2001) Plasma membrane rafts play a critical role in HIV-1 assembly and release. Proc Natl Acad Sci U S A 98:13925–13930
Manes S, del Real G, Lacalle RA et al (2000) Membrane raft microdomains mediate lateral assemblies required for HIV-1 infection. EMBO Rep 1:190–196
Koseki M, Hirano K-i, Masuda D et al (2007) Increased lipid rafts and accelerated lipopolysaccharide-induced tumor necrosis factor-{alpha} secretion in Abca1-deficient macrophages. J Lipid Res 48:299–306
Landry YD, Denis M, Nandi S et al (2006) ATP-binding cassette transporter A1 expression disrupts raft membrane microdomains through its ATPase-related functions. J Biol Chem 281:36091–36101
Cui HL, Grant A, Mukhamedova N et al (2012) HIV-1 Nef mobilizes lipid rafts in macrophages through a pathway that competes with ABCA1-dependent cholesterol efflux. J Lipid Res 53:696–708
Brichacek B, Darwish C, Popratiloff A et al (2014) HIV-1 Infection of macrophages induces retention of cholesterol transporter ABCA1 in the endoplasmic reticulum. AIDS Res Hum Retroviruses 30:947
Jennelle L, Hunegnaw R, Dubrovsky L et al (2014) HIV-1 protein nef inhibits activity of ATP binding cassette transporter A1 by targeting endoplasmic reticulum chaperone calnexin. J Biol Chem 289:28870
Asztalos BF, Mujawar Z, Morrow MP et al (2010) Circulating Nef induces dyslipidemia in simian immunodeficiency virus-infected macaques by suppressing cholesterol efflux. J Infect Dis 202:614–623
Cui HL, Ditiatkovski M, Kesani R et al (2014) HIV protein Nef causes dyslipidemia and formation of foam cells in mouse models of atherosclerosis. FASEB J 28:2828–2839
Crowe SM, Westhorpe CL, Mukhamedova N et al (2010) The macrophage: the intersection between HIV infection and atherosclerosis. J Leukoc Biol 87:589–598
Neufeld EB, Remaley AT, Demosky SJ et al (2001) Cellular localization and trafficking of the human ABCA1 transporter. J Biol Chem 276:27584–27590
Acknowledgments
This study was supported by NIH grants R01HL093818, R01HL101274, R21AI108533, P30AI087714, P30HD040677, S10OD010710, 1S10RR025565.
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Mukhamedova, N., Brichacek, B., Darwish, C., Popratiloff, A., Sviridov, D., Bukrinsky, M. (2016). Analysis of ABCA1 and Cholesterol Efflux in HIV-Infected Cells. In: Prasad, V., Kalpana, G. (eds) HIV Protocols. Methods in Molecular Biology, vol 1354. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3046-3_19
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DOI: https://doi.org/10.1007/978-1-4939-3046-3_19
Publisher Name: Humana Press, New York, NY
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