Retroviral cDNA Integration: Mechanism, Applications and Inhibition

Hansen, Mark; Carteau, Sandrine; Hoffmann, Christopher M.; Li, Ling; Bushman, Frederic D.

doi:10.1007/978-1-4899-1739-3_3

Cited by 33 publications

(30 citation statements)

References 203 publications

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“…28 The unintegrated retroviral cDNA can also follow a nonproductive pathway leading to loss of the viral genome, in which circularization takes place by ligation of the LTR sequences (2-LTR circle), homologous recombination between LTRs (a single LTR circle), or integration of the viral cDNA into itself, yielding internally rearranged circular forms. 29 Over a long-term observation period, we observed a rapid decrease in GFP expression in cells transduced with the MLV vector (Figures 4 and 5).…”

Section: Discussionmentioning

confidence: 99%

Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector

et al. 2003

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Section: Discussionmentioning

confidence: 99%

Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector

et al. 2003

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“…This integration process is catalyzed by the viral enzyme integrase (for reviews see Brown, 1997;Hansen et al, 1998). Integrase (IN) can be split by partial proteolysis in three functional domains that are amenable to structure elucidation by X-ray crystallography and high-resolution two-and three-dimensional NMR techniques.…”

Section: Introductionmentioning

confidence: 99%

X-ray absorption spectroscopic studies of zinc in the N-terminal domain of HIV-2 integrase and model compounds

et al. 2002

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X-ray absorption spectroscopy (XAS), including extended X-ray absorption fine structure (EXAFS) and X-ray absorption near-edge structure (XANES) analysis, has been carried out at the ZnKedge of the N-terminal part of the integrase protein of the human immunodeficiency virus, type 2 (HIV-2), and of some zinc coordination compounds. In the presence of excess β-mercaptoethanol, which was present in the NMR structure elucidation of the protein [Eijkelenboomet al.(1997),Curr. Biol.7, 739–746; (2000),J. Biomol. NMR,18, 119–28], the protein spectrum was nearly identical to that recorded in its absence. Comparison of the XANES of the protein with that of model compounds and literature data permits the conclusion that the Zn ion is four-coordinated. The major shell of the EXAFS provides evidence for a mixed (N or O as well as S) coordination sphere, while the minor shells indicate imidazole coordination. Our approach to the analysis of the EXAFS, including quantification of the imidazole by multiple scattering simulations withEXCURV92, was validated on the model compounds. An important result is that with multiple scattering simulations using restraints on the parameters of the imidazole rings the number of imidazoles and their orientation could be determined. The integrase spectra can be fitted with two sulfur ligands at 2.26 Å (Debye–Waller-type factor 0.009 Å2) and two imidazole ligands with the N atoms at 1.99 Å (Debye–Waller-type factor 0.005 Å2). The XAS-derived geometry is fully consistent with that found in the NMR structure determination and, allowing for the volume contraction due to the temperature difference between the experiments, justifies the restraints applied in the structure calculation (Zn—S and Zn—N distances of 2.3 Å and 2.0 Å, respectively).

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“…Retroviral integration, the process that stably insert the DNA copy of the viral genomic RNA into the host cell genome, is an essential step for productive infection (22,26,30,43,45).…”

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Styrylquinolines, Integrase Inhibitors Acting Prior to Integration: a New Mechanism of Action for Anti-Integrase Agents

Bonnenfant

Thomas

Vita

et al. 2004

J Virol

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We have previously shown that styrylquinolines (SQLs) are integrase inhibitors in vitro. They compete with the long terminal repeat substrate for integrase. Here, we describe the cellular mode of action of these molecules. We show that SQLs do not interfere with virus entry. In fact, concentrations of up to 20 times the 50% inhibitory concentration did not inhibit cell-to-cell fusion or affect the interaction between GP120 and CD4 in vitro. Moreover, the pseudotype of the retrovirus envelope did not affect drug activity. Quantitative reverse transcription PCR experiments showed that SQLs do not inhibit the entry of the genomic RNA. In contrast, the treatment of human immunodeficiency virus type 1-infected cells with SQLs reduced the amount of the late cDNA, suggesting for the first time that integrase targeting molecules may affect the accumulation of DNA during reverse transcription. The cellular target of SQLs was confirmed by the appearance of mutations in the integrase gene when viruses were grown in the presence of increasing concentrations of SQLs. Finally, these mutations led to SQL-resistant viruses when introduced into the wild-type sequence. In contrast, SQLs were fully active against reverse transcriptase inhibitor-and diketo acid-resistant viruses, positioning SQLs as a second group of anti-integrase compounds.

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Retroviral cDNA Integration: Mechanism, Applications and Inhibition

Cited by 33 publications

References 203 publications

Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector

Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector

X-ray absorption spectroscopic studies of zinc in the N-terminal domain of HIV-2 integrase and model compounds

Styrylquinolines, Integrase Inhibitors Acting Prior to Integration: a New Mechanism of Action for Anti-Integrase Agents

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