Circularization of human immunodeficiency virus type 1 DNA in vitro

Farnet, Chris M.; Haseltine, William A.

doi:10.1128/jvi.65.12.6942-6952.1991

Cited by 133 publications

(61 citation statements)

References 32 publications

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“…The first was 2LTR HIV DNA circles that primarily forms within the nucleus and increases if HIV integration is blocked. 48,49 A decrease in 2LTR DNA post-HIV fusion would support a block in nuclear import, whilst an increase would support a block in integration. Secondly, we detected genomically integrated HIV DNA using Alu-HIV PCR [50][51][52] to confirm if HIV has successfully integrated into the human genome.…”

Section: Resolution Of Hiv Post-entry Blocks Mediated By Dynamin Inmentioning

confidence: 99%

HIV infection is influenced by dynamin at 3 independent points in the viral life cycle

Aggarwal

Hitchen

Ootes

et al. 2017

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CD4 T cells are important cellular targets for HIV-1, yet the primary site of HIV fusion remains unresolved. Candidate fusion sites are either the plasma membrane or from within endosomes. One area of investigation compounding the controversy of this field, is the role of the protein dynamin in the HIV life cycle. To understand the role of dynamin in primary CD4 T cells we combined dynamin inhibition with a series of complementary assays based on single particle tracking, HIV fusion, detection of HIV DNA products and active viral transcription. We identify 3 levels of dynamin influence on the HIV life cycle. Firstly, dynamin influences productive infection by preventing cell cycle progression. Secondly, dynamin influences endocytosis rates and increases the probability of endosomal fusion. Finally, we provide evidence in resting CD4 T cells that dynamin directly regulates the HIV fusion reaction at the plasma membrane. We confirm this latter observation using 2 divergent dynamin modulating compounds, one that enhances dynamin conformations associated with dynamin ring formation (ryngo-1-23) and the other that preferentially targets dynamin conformations that appear in helices (dyngo-4a). This in-depth understanding of dynamin's roles in HIV infection clarifies recent controversies and furthermore provides evidence for dynamin regulation specifically in the HIV fusion reaction.

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Section: Resolution Of Hiv Post-entry Blocks Mediated By Dynamin Inmentioning

confidence: 99%

HIV infection is influenced by dynamin at 3 independent points in the viral life cycle

Aggarwal

Hitchen

Ootes

et al. 2017

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“…To determine if ADAM10 is required for proviral DNA integration, genomic and small non-genomic cDNA was isolated from cells at various time points post-infection. If HIV-1 cDNA enters the nucleus but does not integrate into the host cell chromosome, then the viral cDNA circularizes to form a 2-LTR circle [31,32], which can be quantified using real-time PCR. Integration is quantified by using one primer directed against HIV-1 and another primer directed against Alu, a common repetitive sequence found in the human genome.…”

Section: Adam10 Activity In Hiv-1 Replication Precedes Viral Integrationmentioning

confidence: 99%

A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication

Friedrich

Murray²,

et al. 2011

Retrovirology

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BackgroundGene trap insertional mutagenesis was used as a high-throughput approach to discover cellular genes participating in viral infection by screening libraries of cells selected for survival from lytic infection with a variety of viruses. Cells harboring a disrupted ADAM10 (A Disintegrin and Metalloprotease 10) allele survived reovirus infection, and subsequently ADAM10 was shown by RNA interference to be important for replication of HIV-1.ResultsSilencing ADAM10 expression with small interfering RNA (siRNA) 48 hours before infection significantly inhibited HIV-1 replication in primary human monocyte-derived macrophages and in CD4+ cell lines. In agreement, ADAM10 over-expression significantly increased HIV-1 replication. ADAM10 down-regulation did not inhibit viral reverse transcription, indicating that viral entry and uncoating are also independent of ADAM10 expression. Integration of HIV-1 cDNA was reduced in ADAM10 down-regulated cells; however, concomitant 2-LTR circle formation was not detected, suggesting that HIV-1 does not enter the nucleus. Further, ADAM10 silencing inhibited downstream reporter gene expression and viral protein translation. Interestingly, we found that while the metalloprotease domain of ADAM10 is not required for HIV-1 replication, ADAM15 and γ-secretase (which proteolytically release the extracellular and intracellular domains of ADAM10 from the plasma membrane, respectively) do support productive infection.ConclusionsWe propose that ADAM10 facilitates replication at the level of nuclear trafficking. Collectively, our data support a model whereby ADAM10 is cleaved by ADAM15 and γ-secretase and that the ADAM10 intracellular domain directly facilitates HIV-1 nuclear trafficking. Thus, ADAM10 represents a novel cellular target class for development of antiretroviral drugs.

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“…The circularized double-stranded linear DNA, contains either a single copy or a tandem double copy of the long terminal repeats (1-LTR and 2-LTR forms respectively) [1,7,8]. In particular, 1-LTR circles can be produced by autointegration reaction, leading to rearranged circular forms [9,10], reverse transcription intermediates failing to complete the reverse transcription process [11], or homologous recombination between the two LTR [10]. In addition, reverse transcribed linear DNA can also be a substrate for the host cell nonhomologous DNA end joining (NHEJ) pathway, which normally repairs cellular doublestranded breaks by end-ligation, yielding the 2-LTR circular forms.…”

Section: Introductionmentioning

confidence: 99%

Retroviral E-DNA: persistence and gene expression in nondividing immune cells

Cara

Klotman

2006

Journal of Leukocyte Biology

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Following retroviral infection of cells, not only is the proviral DNA integrated into the host genome, but there is also an accumulation of unintegrated extrachromosomal DNA (E-DNA), both linear and circular. Although the integrated DNA is responsible for the production of viral proteins and new viral progeny, the role of E-DNA has remained uncertain. Several reports have shown that E-DNA is transcriptionally active producing both RNA, as well as viral proteins and that circular E-DNA can persist in nondividing cells, raising questions regarding the potential consequences of this reservoir. Furthermore, integrase inhibitors, presently in clinical trials, shifts the balance of proviral DNA to the E-DNA form. This review is focused on recent work in this field with an emphasis on exploring the potential role of E-DNA in both pathogenesis of retroviral infections, especially HIV-1, and as a tool to deliver and express genes.

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Circularization of human immunodeficiency virus type 1 DNA in vitro

Cited by 133 publications

References 32 publications

HIV infection is influenced by dynamin at 3 independent points in the viral life cycle

HIV infection is influenced by dynamin at 3 independent points in the viral life cycle

A Functional Role for ADAM10 in Human Immunodeficiency Virus Type-1 Replication

Retroviral E-DNA: persistence and gene expression in nondividing immune cells

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