Titlebook: Human Retroviruses; Methods and Protocol Elisa Vicenzi,Guido Poli Book 2014 Springer Science+Business Media, LLC 2014 Accessory genes.HIV l

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The Fate of HIV-1 Capsid: A Biochemical Assay for HIV-1 Uncoatingeld forward. Here we describe an uncoating assay that currently represents the state-of-the-art biochemical procedure for monitoring uncoating and core stability during infection. This assay is based on the biochemical separation of soluble capsid protein from particulate capsid cores and provides i

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The Cyclosporin A Washout Assay to Detect HIV-1 Uncoating in Infected Cellsx. Although uncoating is required for HIV-1 replication, many questions remain about the mechanism of this process as well as its impact on other steps in viral replication. Here we describe a recently developed assay to study the process of uncoating in HIV-1-infected cells. The CsA washout assay i

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Imaging HIV-1 Nuclear Pre-integration Complexeslogy. Here we describe a novel, fluorescence-based method to visualize HIV-1 viral particles within intact nuclei of infected cells. This method allows investigating the localization of pre-integration complexes within the nuclear compartment with respect to the nuclear envelope and the chromatin te

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HIV-1 Reverse Transcriptiones the single-stranded positive sense RNA genome to synthesize the double-stranded viral DNA. At the same time the RT-associated RNaseH activity degrades the genomic RNA template, which has just been copied. The viral nucleocapsid protein NCp7 is an obligatory partner of RT, chaperoning synthesis of

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RNase H: Specificity, Mechanisms of Action, and Antiviral Targety in RNA–DNA hybrids. The RNase H ribonuclease is essential in the retroviral life cycle, since it generates and removes primers needed by the Reverse Transcriptase (RT) for initiation of DNA synthesis. Retroviruses lacking RNase H activity are noninfectious. Despite its importance, RNase H is the o

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HIV-1 Chromatin, Transcription, and the Regulatory Protein Tatcriptional regulation. Therefore, characterization of the chromatin changes that occur in the viral promoter region in response to different cellular stimuli or drug treatments represents an important aspect of our understanding of HIV-1 transcription. Moreover, the viral transactivator Tat protein

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HIV-1 Rev Function and RNA Nuclear-Cytoplasmic Export cellular mechanism that block nuclear-cytoplasmic export of incompletely processed mRNAs. Evaluating the function of these viral factors can be done at multiple levels: examining the functional consequence of Rev/Rex on viral gene expression, monitoring the movement of these proteins between the nu