Abstract
BackgroundThe human immunodeficiency virus type 1 (HIV-1) genome (~9 kb RNA) is flanked by two long terminal repeats (LTR) promoter regions with nine open reading frames, which encode Gag, Pol and Env polyproteins, four accessory proteins (Vpu, Vif, Vpr, Nef) and two regulatory proteins (Rev, Tat). In this study, we carried out a genome-wide and functional analysis of the HIV-1 genome in fission yeast (Schizosaccharomyces pombe).ResultsEach one of the HIV-1 genes was cloned and expressed individually in fission yeast. Subcellular localization of each viral protein was first examined. The effect of protein expression on cellular proliferation and colony formations, an indication of cytotoxicity, were observed. Overall, there is a general correlation of subcellular localization of each viral protein between fission yeast and mammalian cells. Three viral proteins, viral protein R (Vpr), protease (PR) and regulator of expression of viral protein (Rev), were found to inhibit cellular proliferation. Rev was chosen for further analysis in fission yeast and mammalian cells. Consistent with the observation in fission yeast, expression of HIV-1 rev gene also caused growth retardation in mammalian cells. However, the observed growth delay was neither due to the cytotoxic effect nor due to alterations in cell cycling. Mechanistic testing of the Rev effect suggests it triggers transient induction of cellular oxidative stress.ConclusionsSome of the behavioral and functional similarities of Rev between fission yeast and mammalian cells suggest fission yeast might be a useful model system for further studies of molecular functions of Rev and other HIV-1 viral proteins.
Highlights
The human immunodeficiency virus type 1 (HIV-1) genome (~9 kb RNA) is flanked by two long terminal repeats (LTR) promoter regions with nine open reading frames, which encode Gag, Pol and Env polyproteins, four accessory proteins (Vpu, Vif, viral protein R (Vpr), Nef ) and two regulatory proteins (Rev, Tat)
Subcellular localizations of HIV‐1 proteins in fission yeast In order to determine the subcellular localization of HIV-1 proteins in fission yeast, SP223 cells were transformed with a fission yeast expression pYZ3N plasmid producing each of the HIV-1 viral protein sequences in fusion with an N-terminal GFP [48]
The fission yeast strains containing different viral proteins expressing plasmids were inoculated into liquid selective medium and the protein expression was induced following the removal of thiamine from the growth medium as described previously [34, 49]
Summary
The human immunodeficiency virus type 1 (HIV-1) genome (~9 kb RNA) is flanked by two long terminal repeats (LTR) promoter regions with nine open reading frames, which encode Gag, Pol and Env polyproteins, four accessory proteins (Vpu, Vif, Vpr, Nef ) and two regulatory proteins (Rev, Tat). The human immunodeficiency virus type 1 (HIV-1), like other retroviruses, is made up of an RNA encoded genome of approximately 9.7 kilobases (kb) Both ends of the RNA genome are flanked by a long terminal repeat (LTR) promoter region (Fig. 1). P24 (Capsid domain, CA) protein forms a shell surrounding the viral RNA genome and core-associated proteins in mature virion It plays various roles including incorporation of the Gag-Pol precursor into virions during viral assembly [8], recruitment of the
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